IL-7 Binding Proteins and Their Use in Medical Therapy

ABSTRACT

Provided herein are interleukin 7 (IL-7) binding proteins, pharmaceutical compositions and their use in the treatment or prevention of a disease or condition.

FIELD OF THE DISCLOSURE

The present disclosure relates to interleukin 7 (IL-7) binding proteins.The disclosure also concerns methods of treating diseases or disorderswith IL-7 binding proteins, uses of IL-7 binding proteins,pharmaceutical compositions comprising IL-7 binding proteins and methodsof their manufacture. Other aspects of the disclosure will be apparentfrom the description below.

BACKGROUND TO THE DISCLOSURE

Both altered autoimmune T cell responses and dysfunction of theregulatory network of the immune system play an important role in humanautoimmune pathologies, such as multiple sclerosis (MS) and rheumatoidarthritis (Kuchroo et al., (2002) Annu. Rev. Immunol. 20:101-123;Sospedra and Martin (2005) Annu. Rev. Immunol. 23: 683-747; Toh andMiossec (2007) Curr. Opin. Rheumatol. 19:284-288).

Although the etiology and pathogenesis of MS remain unknown, it isgenerally considered an autoimmune pathology in which autoreactive Tcells of pathogenic potential, such as T_(H)1 and T_(H)17 cells, arethought to play an important role. There is evidence that these effectorT cells are activated in vivo during the disease process and areattributable to the central nervous system (CNS) inflammation. There isalso evidence that these T cells mediate destruction ofmyelin-expressing cells in lesions of experimental autoimmuneencephalomyelitis (EAE) and MS during the active phase of the disease.On the other hand, regulatory T cells (T_(reg)) that normally keeppathogenic T_(H)1 and T_(H)17 cells in check are deficient in patientswith MS, further tilting the immune system toward a pro-inflammatorystate.

IL-7 and IL-7 receptor (IL-7R) are known to play an important role in Tcell and B cell development and homeostasis mainly in a thymicenvironment. In humans, IL-7 is important for T cell development in thethymus and for survival of memory and naïve T cells in the periphery.Indeed, thymic stromal cells, fetal thymus, and bone marrow are sites ofIL-7 production.

IL-7 is a four-helix bundle, displaying the conserved up-up-down-downcytokine architecture. Based on the work of McElroy et al. (Structure,2009, 17:54-65), the helices of IL-7, starting with that closest to theN-terminus, stretch from amino acids 10-26 (Helix A), amino acids 53-63(Helix B), amino acids 74-91 (Helix C) and amino acids 128-146 (HelixD). A mini-helix is believed to exist between Helix A and Helix B, andstretches from amino acids 40-44. For the purposes of the disclosure,Helices A, B, C and D are referred to as Helices 1, 3, 4 and 5, and themini-helix is referred to as Helix 2. The loops interconnecting thehelices therefore stretch from amino acids 27-39 (“Loop 1” between Helix1 and 2), 45 to 52 (“Loop 2” between Helix 2 and 3), 64-73 (“Loop 3”between Helix 3 and 4), and 92-127 (“Loop 4” between Helix 4 and 5). The“N-terminal loop” runs from amino acids 1-9, and the “C-terminal loop”from amino acids 147-152.

The IL-7 receptor consists of two subunits, CD127 and a common chain(gamma chain or γc) which is shared by receptors of IL-2, IL-4, IL-9,IL-15, and IL-21. CD127 is also known as IL-7 receptor alpha (IL-7Rα)and p90 IL-7R. Human CD127 (Swiss Prot accession number P16871) has atotal of 459 amino acids (20 signal sequence). It comprises a 219 aminoacid extra cellular region, a 25 amino acid transmembrane region and a195 amino acid intracellular region. Both IL-7Rα and γc are required forsignal transduction. The crystal structure of an IL-7/IL-7Rα ECD(extracellular domain) complex has been resolved (McElroy et al.,supra), but to date there is limited information about the interactionbetween IL-7 and the functional IL-7Rα/γc receptor complex. The residuesof IL-7 importance in this interaction have not been determined,although predictions generally implicate Helix A (Helix 1, herein) andHelix D (Helix 5, herein) in the binding of IL-7 to γc.

Binding of IL-7 to the IL-7R complex activates multiple signalingpathways including the activation of JAK kinases 1 and 3 leading to thephosphorylation and activation of STAT5. This pathway is crucial to thesurvival of thymic developing T cell precursors because STAT5 activationis required in the induction of the anti-apoptotic protein Bcl-2 and theprevention of the pro-apoptotic protein Bax entry into themitochondrion. Another IL-7R mediated pathway is the activation of PI3kinase, resulting in the phosphorylation of the pro-apoptotic proteinBAD (BCL2 associated agonist of cell death) and its cytoplasm retention.

Pathogenic CD4⁺ T cells in multiple sclerosis (MS) are largely fromT_(H)1 and T_(H)17 subsets which is reflected by the pro-inflammatorycytokine production from these cells—IFN-γ, IL-17 and GM-CSF etc. Thesecytokines contribute to blood-brain-barrier dysfunction, inflammationand activation of resident astrocytes and microglia cells. IL-7 caninduce Th₁ and Th₁₇ phenotypes upon antigen stimulation and effectormemory T cells rely on IL-7 for survival and proliferation. Thepathogenic autoreactive T cells in MS are maintained by a pool of memoryT cells and IL-7 is key for their development, proliferation andactivation. Accordingly, it is desirable to develop antagonists of theIL-7/IL-7R pathway. Such antagonists may be therapeutically useful inthe treatment of MS and other inflammatory and/or autoimmune diseasesand disorders, particularly those in which raised T_(H)1 and/or T_(H)17cells have been observed.

SUMMARY OF THE DISCLOSURE

Disclosed herein are IL-7 binding proteins, for example an IL-7 bindingantibody or an antigen binding fragment thereof.

In one aspect of the invention, there is provided an IL-7 bindingprotein that binds to one or more amino acid residue within the aminoacid sequence set forth in SEQ ID NO:12 of human IL-7. In oneembodiment, the IL-7 binding protein protects residues 67 to 81 (SEQ IDNO:12) of IL-7 from deuterium exchange in HDX-MS analysis. In a furtherembodiment, the IL-7 binding protein protects residues 67 to 80 (SEQ IDNO:16) of IL-7 from deuterium exchange in HDX-MS analysis.

In another aspect of the invention, there is provided an IL-7 bindingprotein that binds to human IL-7 adjacent an IL-7Rα binding site, with aKD of 100 nM or less as measured by surface plasmon resonance assay. Insome embodiments, the IL-7 binding protein inhibits IL-7 binding toIL-7R as measured in an in vitro competitive binding assay as determinedusing a surface plasmon resonance assay. In other embodiments, the IL-7binding protein further comprises one or more of CDRH1 as set out in SEQID NO:6, CDRH2 as set out in SEQ ID NO:7, CDRH3 as set out in SEQ IDNO:8, CDRL1 as set out in SEQ ID NO:9, CDRL2 as set out in SEQ ID NO:10and/or CDRL3 as set out in SEQ ID NO:11.

In a further aspect of the invention, the IL-7 binding protein comprisesCDRH1 as set out in SEQ ID NO:6, CDRH2 as set out in SEQ ID NO:7, CDRH3as set out in SEQ ID NO:8, CDRL1 as set out in SEQ ID NO:9, CDRL2 as setout in SEQ ID NO:10 and CDRL3 as set out in SEQ ID NO:11. In someembodiments, the IL-7 binding protein comprises a V_(H) domain having atleast 90% identity to the amino acid sequence set out in SEQ ID NO:4and/or a VL domain having at least 90% identity to the amino acidsequence as set out in SEQ ID NO:5. In another embodiment, the IL-7binding protein comprises a V_(H) domain as set out in SEQ ID NO:4 and aVL domain as set out in SEQ ID NO:5. In some embodiments, the IL-7binding protein comprises a constant region such that the IL-7 bindingprotein has reduced ADCC and/or complement activation or effectorfunctionality. In some embodiments, the IL-7 binding protein comprises aheavy chain Fc domain having an alanine residue at position 235 andposition 237 according to EU numbering. In other embodiments, the IL-7binding protein comprises a scaffold selected from the group consistingof human IgG1 isotype and human IgG4 isotype. In one embodiment, theIL-7 binding protein is of human IgG1 isotype. In another embodiment,the IL-7 binding protein is a monoclonal antibody. In some embodiments,the monoclonal antibody is human, humanized or chimeric. In otherembodiments, the IL-7 binding protein comprises a heavy chain having atleast 90% identity to the amino acid sequence set out in SEQ ID NO:2 anda light chain having at least 90% identity to the amino acid sequence asset out in SEQ ID NO:3. In one embodiment, the IL-7 binding proteincomprises a heavy chain as set out in SEQ ID NO:2 and a light chain asset out in SEQ ID NO:3. In some embodiments, the IL-7 binding proteinbinds to IL-7 with a KD of 50 nM or less, 10 nM or less, 1 nM or less,or 0.1 nM or less. In another embodiment, the IL-7 binding protein bindsto IL-7 and (i) inhibits IL-7 dependent IFN-γ or IL-10 secretion fromperipheral blood mononuclear cells with an IC₅₀ of 1 nM or less, and/or(ii) inhibits IL-7 dependent STAT5 phosphorylation in CD4+ T cells withan IC₅₀ of 1 nM or less. In some embodiments, the IL-7 binding proteininhibits signaling, activation, cytokine production and proliferation ofCD4⁺ T cells and/or CD8⁺ T cells.

In one aspect of the invention, there is provided a nucleic acidencoding an IL-7 binding protein. In another aspect of the invention,there is provided an expression vector comprising a nucleic acidsequence encoding the IL-7 binding protein. In a further aspect of theinvention, there is provided a host cell comprising the nucleic acidsequence encoding the IL-7 binding protein or the expression vectorcomprising the nucleic acid sequence encoding the IL-7 binding protein.In another aspect of the invention, there is provided an IL-7 bindingprotein expressed by the host cell.

In another aspect of the invention, there is provided a method of makingan IL-7 binding protein, the method comprising maintaining a host cellcomprising a nucleic acid sequence encoding the IL-7 binding protein orthe expression vector comprising the nucleic acid sequence encoding theIL-7 binding protein, in a medium to produce the IL-7 binding proteinand isolating or purifying the IL-7 binding protein produced by the hostcell.

In a further aspect of the invention, there is provided an IL-7 bindingprotein that competes for binding to IL-7 with the IL-7 binding proteindisclosed herein.

In yet another aspect of the invention, there is provided apharmaceutical composition, comprising a pharmaceutically-acceptablecarrier or excipient and an IL-7 binding protein that exhibits bindingfor IL-7 at an epitope comprising at least 5 contiguous amino acids of asequence set out in SEQ ID NO:12 or SEQ ID NO:16. In a further aspect ofthe invention, there is provided a pharmaceutical composition comprisingthe IL-7 binding protein and a pharmaceutically acceptable carrier orexcipient.

In another aspect of the invention, there is provided an IL-7 bindingprotein for use in therapy. In one aspect of the invention, there isprovided a method for the treatment of an autoimmune and/or inflammatorycondition in a subject in need thereof, comprising administering to thesubject a therapeutically effective amount of the IL-7 binding proteinor the pharmaceutical composition. In one aspect of the invention, thereis provided an IL-7 binding protein for use in the treatment of anautoimmune and/or inflammatory condition. In another aspect of theinvention, there is provided use of an IL-7 binding protein in themanufacture of a medicament for treatment of an autoimmune and/orinflammatory condition. In some embodiments, the autoimmune and/orinflammatory condition is multiple sclerosis, Sjögren's syndrome,rheumatoid arthritis, Crohn's disease, ulcerative colitis or lupuserythematosus. In some embodiments, the multiple sclerosis is clinicallyisolated syndrome, relapsing remitting, primary progressive or secondaryprogressive.

In a further aspect of the invention, there is provided a kit comprisingthe IL-7 binding protein and instructions for use.

In one aspect of the invention, there is provided an IL-7 bindingprotein comprising CDRH1 as set out in SEQ ID NO:26, CDRH2 as set out inSEQ ID NO:27, CDRH3 as set out in SEQ ID NO:28, CDRL1 as set out in SEQID NO:29, CDRL2 as set out in SEQ ID NO:30 and CDRL3 as set out in SEQID NO:31. In some embodiments, the IL-7 binding protein comprises aV_(H) domain having at least 90% identity to the amino acid sequence setout in SEQ ID NO:25 and/or a VL domain having at least 90% identity tothe amino acid sequence as set out in SEQ ID NO:24. In anotherembodiment, the IL-7 binding protein comprises a V_(H) domain as set outin SEQ ID NO:25 and a VL domain as set out in SEQ ID NO:24.

In one aspect of the invention, there is provided an IL-7 bindingprotein comprising CDRH1 as set out in SEQ ID NO:34, CDRH2 as set out inSEQ ID NO:35, CDRH3 as set out in SEQ ID NO:36, CDRL1 as set out in SEQID NO:37, CDRL2 as set out in SEQ ID NO:38 and CDRL3 as set out in SEQID NO:39. In some embodiments, the IL-7 binding protein comprises aV_(H) domain having at least 90% identity to the amino acid sequence setout in SEQ ID NO:33 and/or a VL domain having at least 90% identity tothe amino acid sequence as set out in SEQ ID NO:32. In anotherembodiment, the IL-7 binding protein comprises a V_(H) domain as set outin SEQ ID NO:33 and a VL domain as set out in SEQ ID NO:32.

In one aspect of the invention, there is provided an IL-7 bindingprotein comprising CDRH1 as set out in SEQ ID NO:42, CDRH2 as set out inSEQ ID NO:43, CDRH3 as set out in SEQ ID NO:44, CDRL1 as set out in SEQID NO:45, CDRL2 as set out in SEQ ID NO:46 and CDRL3 as set out in SEQID NO:47. In some embodiments, the IL-7 binding protein comprises aV_(H) domain having at least 90% identity to the amino acid sequence setout in SEQ ID NO:41 and/or a VL domain having at least 90% identity tothe amino acid sequence as set out in SEQ ID NO:40. In anotherembodiment, the IL-7 binding protein comprises a V_(H) domain as set outin SEQ ID NO:41 and a VL domain as set out in SEQ ID NO:40.

In some embodiments, the IL-7 binding protein or an antigen bindingfragment thereof binds to one or more residues within the amino acidsequence set forth in SEQ ID NO:12 of human IL-7. In some embodiments,the IL-7 binding protein protects residues 67 to 81 (SEQ ID NO:12) ofIL-7 from deuterium exchange in HDX-MS analysis. In some embodiments,the IL-7 binding protein or an antigen binding fragment thereof binds toone or more residues within the amino acid sequence set forth in SEQ IDNO:16 of human IL-7. In some embodiments, the IL-7 binding proteinprotects residues 67 to 80 (SEQ ID NO:16) of IL-7 from deuteriumexchange in HDX-MS analysis. In some embodiments, the IL-7 bindingprotein binds to an epitope comprising the amino acid sequence set forthin SEQ ID NO:12. In some embodiments, the IL-7 binding protein binds toan epitope comprising at least 50%, 60%, 70%, 80%, 90% or 95% identityto the amino acid sequence set out in SEQ ID NO:12. In some embodiments,the IL-7 binding protein comprises a variable region light chain havingat least 80% identity to the amino acid sequence set out in SEQ ID NO:5.In some embodiments, the IL-7 binding protein comprises at least one of(a) a heavy chain CDR1 having at least 80% identity to the amino acidsequence set out in SEQ ID NO:6, (b) a heavy chain CDR2 having at least80% identity to the amino acid sequence set out in SEQ ID NO:7 or (c) aheavy chain CDR3 having at least 80% identity to the amino acid sequenceset out in SEQ ID NO:8. In some embodiments, the IL-7 binding proteincomprises at least one of (a) a light chain CDR1 having at least 80%identity to the amino acid sequence set out in SEQ ID NO:9, (b) a lightchain CDR2 having at least 80% identity to the amino acid sequence setout in SEQ ID NO:10 or (c) a light chain CDR3 having at least 80%identity to the amino acid sequence set out in SEQ ID NO:11. In someembodiments, the IL-7 binding protein comprises a variable region lightchain having the amino acid sequence set out in SEQ ID NO:5. In someembodiments, the IL-7 binding protein comprises a heavy chain CDR1comprising the amino acid sequence set out in SEQ ID NO:6, a heavy chainCDR2 comprising the amino acid sequence set out in SEQ ID NO:7 and aheavy chain CDR3 comprising the amino acid sequence set out in SEQ IDNO:8. In some embodiments, the IL-7 binding protein comprises a lightchain CDR1 comprising the amino acid sequence set out in SEQ ID NO:9, alight chain CDR2 comprising the amino acid sequence set out in SEQ IDNO:10 and a light chain CDR3 the amino acid sequence set out in SEQ IDNO:11. In some embodiments, the IL-7 binding protein comprises avariable region heavy chain having at least 80% identity to the aminoacid sequence set out in SEQ ID NO:4. In some embodiments, the IL-7binding protein comprises a variable region heavy chain comprising theamino acid sequence set out in SEQ ID NO:4. In some embodiments, theIL-7 binding protein comprises a light chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein comprises a light chain comprisingthe amino acid sequence set out in SEQ ID NO:3. In some embodiments, theIL-7 binding protein comprises a heavy chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:2. In someembodiments, the IL-7 binding protein comprises a heavy chain comprisingthe amino acid sequence set out in SEQ ID NO:2. In some embodiments, theIL-7 binding protein comprises a heavy chain comprising the amino acidsequence set out in SEQ ID NO:2 and a light chain comprising the aminoacid sequence set out in SEQ ID NO:3. In some embodiments, the IL-7binding protein is an antibody or an antigen-binding portion thereof. Insome embodiments, the antibody is a monoclonal antibody. In someembodiments, the monoclonal antibody is an IgG antibody. In someembodiments, the antibody is a IgG1 isotype antibody. In someembodiments, the monoclonal antibody is an IgG4 isotype antibody. Insome embodiments, the IL-7 binding protein comprises a constant regionsuch that the IL-7 binding protein has reduced ADCC and/or complementactivation or effector functionality. In some embodiments, the IL-7binding protein comprises a heavy chain Fc domain having an alanineresidue at position 235 and/or position 237 according to EU numbering.In some embodiments, the IL-7 binding protein is human, humanized orchimeric. In some embodiments, the IL-7 binding protein is humanized. Insome embodiments, the IL-7 binding protein is human. In someembodiments, the IL-7 binding protein binds to and neutralizes IL-7. Insome embodiments, the IL-7 binding protein binds to circulating IL-7. Insome embodiments, the IL-7 binding protein binds to IL-7 and inhibitsIL-7 depending IFN-γ or IL-10 secretion from peripheral bloodmononuclear cells with an IC₅₀ of 1 nM or less. In some embodiments, theIL-7 binding protein is an isolated IL-7 binding protein. In someembodiments, the IL-7 binding protein binds to IL-7 and inhibits IL-7depending STAT5 phosphorylation in CD4+ T cells with an IC₅₀ of 1 nM orless. In some embodiments, the IL-7 binding protein is a reversibledimer. In some embodiments, the IL-7 binding protein is a dimer. In someembodiments, the IL-7 binding protein is a monomer. In some embodiments,the IL-7 binding protein inhibits signaling, activation, cytokineproduction and proliferation of CD4⁺ T cells and/or CD8⁺ T cells. Insome embodiments, the IL-7 binding protein is for use in a therapy.Further disclosed are nucleic acids encoding the IL-7 binding protein.In some embodiments, the nucleic acid comprises a sequence having atleast 80% identity to the nucleic acid sequence set out in SEQ ID NO:13encoding the light chain. In some embodiments, the nucleic acidcomprises the nucleic acid sequence set out in SEQ ID NO:13 encoding thelight chain. In some embodiments, the nucleic acid comprises a sequencehaving at least 80% identity to the nucleic acid sequence set out in SEQID NO:14 encoding the heavy chain. In some embodiments, the nucleic acidcomprises the nucleic acid sequence set out in SEQ ID NO:14 encoding theheavy chain. In some embodiments, the nucleic acid further comprises asequence having at least 80% identity to the nucleic acid sequence setout in SEQ ID NO:15 encoding a signal peptide. In some embodiments,disclosed herein are vectors comprising a nucleic acid. In someembodiments, the vector comprises a promoter functional in a mammaliancell. Also disclosed are host cells comprising the nucleic acid. In someembodiments, the host cell is a CHO cell. Further disclosed herein aremethods for making the IL-7 binding protein comprising maintaining thehost cell in a medium to produce the IL-7 binding protein and isolatingor purifying the IL-7 binding protein produced by the host cell. In someembodiments, disclosed herein are pharmaceutical compositions comprisingthe IL-7 binding protein. In some embodiment, the pharmaceuticalcompositions comprises a pharmaceutically acceptable carrier orexcipient. In some embodiments, the pharmaceutical composition furthercomprises at least one additional therapeutic agent. In someembodiments, the pharmaceutical composition has a pH of 4.5-7.0. In someembodiments, the pharmaceutical composition has a pH of 5.5, 6.0, 6.2 or6.5. In some embodiments, the pharmaceutical composition or the IL-7binding protein can be used in a method for the treatment of anautoimmune and/or inflammatory condition in a subject in need thereof,the method comprising administering to the subject a therapeuticallyeffective amount of the pharmaceutical composition or the IL-7 bindingprotein. In some embodiments, the administering is transarterially,subcutaneously, intradermally, intratumorally, intranodally,intramedullary, intramuscularly, intravenously or intraperitoneally. Insome embodiments, the administering is subcutaneously. In someembodiments, the therapeutically effective amount is at least about 0.1mg/kg, 0.5 mg/kg, 1 mg/kg, 3 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg, 30mg/kg, or 40 mg/kg. In some embodiments, the therapeutically effectiveamount is administered to the subject at least about twice every day oronce every 1-60 days. In some embodiments, the therapeutically effectiveamount is administered to the subject once every 4 weeks. In someembodiments, the autoimmune and/or inflammatory condition is Sjögren'ssyndrome. In some embodiments, the autoimmune and/or inflammatorycondition is rheumatoid arthritis. In some embodiments, the autoimmuneand/or inflammatory condition is inflammatory bowel disease (IBD). Insome embodiments, the autoimmune and/or inflammatory condition ismultiple sclerosis. In some embodiments, the multiple sclerosis isclinically isolated syndrome, relapsing-remitting, primary progressiveor secondary progressive. In some embodiments, the autoimmune and/orinflammatory condition is Crohn's disease. In some embodiments, theautoimmune and/or inflammatory condition is ulcerative colitis. In someembodiments, the autoimmune and/or inflammatory condition is lupuserythematosus. In some embodiments, the autoimmune and/or inflammatorycondition is diabetes, for example type I diabetes. In some embodiments,the IL-7 binding protein is used in the manufacture of a medicament forthe treatment of the autoimmune and/or inflammatory condition. In someembodiments, compounds comprising the IL-7 binding protein are for usein the treatment of the autoimmune and/or inflammatory condition. Insome embodiments, the IL-7 binding protein is used for diagnosis of adisease or condition. In some embodiments, the IL-7 binding protein isused for diagnosis of the autoimmune and/or inflammatory condition. Insome embodiments, the IL-7 binding protein is bound to a moiety or anantigenic fragment thereof. In some embodiments, the moiety is IL-7 or afragment thereof. In some embodiments, the IL-7 binding protein isattached directly or indirectly to a solid support. In some embodiments,the IL-7 binding protein is comprised in a kit with instruction for use.In some embodiments, the solid support is comprised in a kit.

Disclosed herein are IL-7 binding proteins or an antigen bindingfragment thereof that binds to human IL-7 with a KD of 100 nM or less.In some embodiments, the IL-7 binding protein binds to human IL-7adjacent IL-7Rα binding site. In some embodiments, the IL-7 bindingprotein inhibits IL-7 binding to IL-7R as measured in an in vitrocompetitive binding assay. In some embodiments, a competitive bindingassay is an immunoassay. In some embodiment, a competitive binding assayis a surface plasmon resonance assay. In some embodiments, a competitivebinding assay is for example ELISA or a radioimmunoassay. In someembodiments, the IL-7 binding protein comprises a variable region lightchain having at least 80% identity to the amino acid sequence set out inSEQ ID NO:5. In some embodiments, the IL-7 binding protein comprises atleast one of (a) a heavy chain CDR1 having at least 80% identity to theamino acid sequence set out in SEQ ID NO:6, (b) a heavy chain CDR2having at least 80% identity to the amino acid sequence set out in SEQID NO:7 or (c) a heavy chain CDR3 having at least 80% identity to theamino acid sequence set out in SEQ ID NO:8. In some embodiments, theIL-7 binding protein comprises at least one of (a) a light chain CDR1having at least 80% identity to the amino acid sequence set out in SEQID NO:9, (b) a light chain CDR2 having at least 80% identity to theamino acid sequence set out in SEQ ID NO:10 or (c) a light chain CDR3having at least 80% identity to the amino acid sequence set out in SEQID NO:11. In some embodiments, the IL-7 binding protein comprises avariable region light chain having the amino acid sequence set out inSEQ ID NO:5. In some embodiments, the IL-7 binding protein comprises aheavy chain CDR1 comprising the amino acid sequence set out in SEQ IDNO:6, a heavy chain CDR2 comprising the amino acid sequence set out inSEQ ID NO:7 and a heavy chain CDR3 comprising the amino acid sequenceset out in SEQ ID NO:8. In some embodiments, the IL-7 binding proteincomprises a light chain CDR1 comprising the amino acid sequence set outin SEQ ID NO:9, a light chain CDR2 comprising the amino acid sequenceset out in SEQ ID NO:10 and a light chain CDR3 the amino acid sequenceset out in SEQ ID NO:11. In some embodiments, the IL-7 binding proteincomprises a variable region heavy chain having at least 80% identity tothe amino acid sequence set out in SEQ ID NO:4. In some embodiments, theIL-7 binding protein comprises a variable region heavy chain comprisingthe amino acid sequence set out in SEQ ID NO:4. In some embodiments, theIL-7 binding protein comprises a light chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein comprises a light chain comprisingthe amino acid sequence set out in SEQ ID NO:3. In some embodiments, theIL-7 binding protein comprises a heavy chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:2. In someembodiments, the IL-7 binding protein comprises a heavy chain comprisingthe amino acid sequence set out in SEQ ID NO:2. In some embodiments, theIL-7 binding protein comprises a heavy chain comprising the amino acidsequence set out in SEQ ID NO:2 and a light chain comprising the aminoacid sequence set out in SEQ ID NO:3. In some embodiments, the IL-7binding protein is an antibody or an antigen-binding portion thereof. Insome embodiments, the antibody is a monoclonal antibody. In someembodiments, the monoclonal antibody is an IgG antibody. In someembodiments, the antibody is a IgG1 isotype antibody. In someembodiments, the monoclonal antibody is an IgG4 isotype antibody. Insome embodiments, the IL-7 binding protein comprises a constant regionsuch that the IL-7 binding protein has reduced ADCC and/or complementactivation or effector functionality. In some embodiments, the IL-7binding protein comprises a heavy chain Fc domain having an alanineresidue at position 235 and/or position 237 according to EU numbering.In some embodiments, the IL-7 binding protein is human, humanized orchimeric. In some embodiments, the IL-7 binding protein is humanized. Insome embodiments, the IL-7 binding protein is human. In someembodiments, the IL-7 binding protein binds to and neutralizes IL-7. Insome embodiments, the IL-7 binding protein binds to circulating IL-7. Insome embodiments, the IL-7 binding protein is an isolated IL-7 bindingprotein. In some embodiments, the IL-7 binding protein binds to IL-7 andinhibits IL-7 depending IFN-γ or IL-10 secretion from peripheral bloodmononuclear cells with an IC₅₀ of 1 nM or less. In some embodiments, theIL-7 binding protein binds to IL-7 and inhibits IL-7 depending STAT5phosphorylation in CD4+ T cells with an IC₅₀ of 1 nM or less. In someembodiments, the IL-7 binding protein inhibits signaling, activation,cytokine production and proliferation of CD4+ T cells and/or CD8+ Tcells. In some embodiments, the IL-7 binding protein is a reversibledimer. In some embodiments, the IL-7 binding protein is a dimer. In someembodiments, the IL-7 binding protein is a monomer. In some embodiments,the IL-7 binding protein is for use in a therapy. Further disclosed arenucleic acids encoding the IL-7 binding protein. In some embodiments,the nucleic acid comprises a sequence having at least 80% identity tothe nucleic acid sequence set out in SEQ ID NO:13 encoding the lightchain. In some embodiments, the nucleic acid comprises the nucleic acidsequence set out in SEQ ID NO:13 encoding the light chain. In someembodiments, the nucleic acid comprises a sequence having at least 80%identity to the nucleic acid sequence set out in SEQ ID NO:14 encodingthe heavy chain. In some embodiments, the nucleic acid comprises thenucleic acid sequence set out in SEQ ID NO:14 encoding the heavy chain.In some embodiments, the nucleic acid further comprises a sequencehaving at least 80% identity to the nucleic acid sequence set out in SEQID NO:15 encoding a signal peptide. In some embodiments, disclosedherein are vectors comprising a nucleic acid. In some embodiments, thevector comprises a promoter functional in a mammalian cell. Alsodisclosed are host cells comprising the nucleic acid. In someembodiments, the host cell is a CHO cell. Further disclosed herein aremethods for making the IL-7 binding protein comprising maintaining thehost cell in a medium to produce the IL-7 binding protein and isolatingor purifying the IL-7 binding protein produced by the host cell. In someembodiments, disclosed herein are pharmaceutical compositions comprisingthe IL-7 binding protein. In some embodiment, the pharmaceuticalcompositions comprises a pharmaceutically acceptable carrier orexcipient. In some embodiments, the pharmaceutical composition furthercomprises at least one additional therapeutic agent. In someembodiments, the pharmaceutical composition has a pH of 4.5-7.0. In someembodiments, the pharmaceutical composition has a pH of 5.5, 6.0, 6.2 or6.5. In some embodiments, the pharmaceutical composition or the IL-7binding protein can be used in a method for the treatment of anautoimmune and/or inflammatory condition in a subject in need thereof,the method comprising administering to the subject a therapeuticallyeffective amount of the pharmaceutical composition or the IL-7 bindingprotein. In some embodiments, the administering is transarterially,subcutaneously, intradermally, intratumorally, intranodally,intramedullary, intramuscularly, intravenously or intraperitoneally. Insome embodiments, the administering is subcutaneously. In someembodiments, the therapeutically effective amount is at least about 0.1mg/kg, 0.5 mg/kg, 1 mg/kg, 3 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg, 30mg/kg, or 40 mg/kg. In some embodiments, the therapeutically effectiveamount is administered to the subject at least about twice every day oronce every 1-60 days. In some embodiments, the therapeutically effectiveamount is administered to the subject once every 4 weeks. In someembodiments, the autoimmune and/or inflammatory condition is Sjögren'ssyndrome. In some embodiments, the autoimmune and/or inflammatorycondition is IBD. In some embodiments, the autoimmune and/orinflammatory condition is rheumatoid arthritis. In some embodiments, theautoimmune and/or inflammatory condition is multiple sclerosis. In someembodiments, the multiple sclerosis is clinically isolated syndrome,relapsing-remitting, primary progressive or secondary progressive. Insome embodiments, the autoimmune and/or inflammatory condition isCrohn's disease. In some embodiments, the autoimmune and/or inflammatorycondition is ulcerative colitis. In some embodiments, the autoimmuneand/or inflammatory condition is lupus erythematosus. In someembodiments, the autoimmune and/or inflammatory condition is diabetes,for example type I diabetes. In some embodiments, the IL-7 bindingprotein is used in the manufacture of a medicament for the treatment ofthe autoimmune and/or inflammatory condition. In some embodiments,provided are compounds comprising the IL-7 binding protein are for usein the treatment of the autoimmune and/or inflammatory condition. Insome embodiments, the IL-7 binding protein is used for diagnosis of adisease or condition. In some embodiments, the IL-7 binding protein isused for diagnosis of the autoimmune and/or inflammatory condition. Insome embodiments, the IL-7 binding protein is bound to a moiety or anantigenic fragment thereof. In some embodiments, the moiety is IL-7 or afragment thereof. In some embodiments, the IL-7 binding protein isattached directly or indirectly to a solid support. In some embodiments,the IL-7 binding protein is comprised in a kit with instruction for use.In some embodiments, the solid support is comprised in a kit.

Disclosed herein are IL-7 binding proteins comprising a variable regionlight chain having at least 95% identity to the amino acid sequence setout in SEQ ID NO:5. In some embodiments, the IL-7 binding proteincomprises at least one of (a) a heavy chain CDR1 having at least 80%identity to the amino acid sequence set out in SEQ ID NO:6, (b) a heavychain CDR2 having at least 80% identity to the amino acid sequence setout in SEQ ID NO:7 or (c) a heavy chain CDR3 having at least 80%identity to the amino acid sequence set out in SEQ ID NO:8. In someembodiments, the IL-7 binding protein comprises a variable region lightchain having the amino acid sequence set out in SEQ ID NO:5. In someembodiments, the IL-7 binding protein comprises a heavy chain CDR1comprising the amino acid sequence set out in SEQ ID NO:6, a heavy chainCDR2 comprising the amino acid sequence set out in SEQ ID NO:7 and aheavy chain CDR3 comprising the amino acid sequence set out in SEQ IDNO:8. In some embodiments, the IL-7 binding protein comprises a lightchain CDR1 comprising the amino acid sequence set out in SEQ ID NO:9, alight chain CDR2 comprising the amino acid sequence set out in SEQ IDNO:10 and a light chain CDR3 the amino acid sequence set out in SEQ IDNO:11. In some embodiments, the IL-7 binding protein comprises avariable region heavy chain having at least 80% identity to the aminoacid sequence set out in SEQ ID NO:4. In some embodiments, the IL-7binding protein comprises a variable region heavy chain comprising theamino acid sequence set out in SEQ ID NO:4. In some embodiments, theIL-7 binding protein comprises a light chain having at least 80%identity to the amino acid set sequence out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein comprises a light chain comprisingthe amino acid sequence set out in SEQ ID NO:3. In some embodiments, theIL-7 binding protein comprises a heavy chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:2. In someembodiments, the IL-7 binding protein comprises a heavy chain comprisingthe amino acid sequence set out in SEQ ID NO:2. In some embodiments, theIL-7 binding protein comprises a heavy chain comprising the amino acidsequence set out in SEQ ID NO:2 and a light chain comprising the aminoacid sequence set out in SEQ ID NO:3. In some embodiments, the IL-7binding protein is an antibody or an antigen-binding portion thereof. Insome embodiments, the antibody is a monoclonal antibody. In someembodiments, the monoclonal antibody is an IgG antibody. In someembodiments, the antibody is a IgG1 isotype antibody. In someembodiments, the monoclonal antibody is an IgG4 isotype antibody. Insome embodiments, the IL-7 binding protein comprises a constant regionsuch that the IL-7 binding protein has reduced ADCC and/or complementactivation or effector functionality. In some embodiments, the IL-7binding protein comprises a heavy chain Fc domain having an alanineresidue at position 235 and/or position 237 according to EU numbering.In some embodiments, the IL-7 binding protein is human, humanized orchimeric. In some embodiments, the IL-7 binding protein is humanized. Insome embodiments, the IL-7 binding protein is human. In someembodiments, the IL-7 binding protein binds to and neutralizes IL-7. Insome embodiments, the IL-7 binding protein binds to native IL-7. In someembodiments, the IL-7 binding protein binds to circulating IL-7. In someembodiments, the IL-7 binding protein binds to IL-7 and inhibits IL-7depending IFN-γ or IL-10 secretion from peripheral blood mononuclearcells with an IC₅₀ of 1 nM or less. In some embodiments, the IL-7binding protein is an isolated IL-7 binding protein. In someembodiments, the IL-7 binding protein binds to IL-7 and inhibits IL-7depending STAT5 phosphorylation in CD4+ T cells with an IC₅₀ of 1 nM orless. In some embodiments, the IL-7 binding protein inhibits signaling,activation, cytokine production and proliferation of CD4+ T cells and/orCD8+ T cells. In some embodiments, the IL-7 binding protein is areversible dimer. In some embodiments, the IL-7 binding protein is adimer. In some embodiments, the IL-7 binding protein is a monomer. Insome embodiments, the IL-7 binding protein is for use in a therapy.Further disclosed are nucleic acids encoding the IL-7 binding protein.In some embodiments, the nucleic acid comprises a sequence having atleast 80% identity to the nucleic acid sequence set out in SEQ ID NO:13encoding the light chain. In some embodiments, the nucleic acidcomprises the nucleic acid sequence set out in SEQ ID NO:13 encoding thelight chain. In some embodiments, the nucleic acid comprises a sequencehaving at least 80% identity to the nucleic acid sequence set out in SEQID NO:14 encoding the heavy chain. In some embodiments, the nucleic acidcomprises the nucleic acid sequence set out in SEQ ID NO:14 encoding theheavy chain. In some embodiments, the nucleic acid further comprises asequence having at least 80% identity to the nucleic acid sequence setout in SEQ ID NO:15 encoding a signal peptide. In some embodiments,disclosed herein are vectors comprising a nucleic acid. In someembodiments, the vector comprises a promoter functional in a mammaliancell. Also disclosed are host cells comprising the nucleic acid. In someembodiments, the host cell is a CHO cell. Further disclosed herein aremethods for making the IL-7 binding protein comprising maintaining thehost cell in a medium to produce the IL-7 binding protein and isolatingor purifying the IL-7 binding protein produced by the host cell. In someembodiments, disclosed herein are pharmaceutical compositions comprisingthe IL-7 binding protein. In some embodiment, the pharmaceuticalcompositions comprises a pharmaceutically acceptable carrier orexcipient. In some embodiments, the pharmaceutical composition furthercomprises at least one additional therapeutic agent. In someembodiments, the pharmaceutical composition has a pH of 4.5-7.0. In someembodiments, the pharmaceutical composition has a pH of 5.5, 6.0, 6.2 or6.5. In some embodiments, the pharmaceutical composition or the IL-7binding protein can be used in a method for the treatment of anautoimmune and/or inflammatory condition in a subject in need thereof,the method comprising administering to the subject a therapeuticallyeffective amount of the pharmaceutical composition or the IL-7 bindingprotein. In some embodiments, the administering is transarterially,subcutaneously, intradermally, intratumorally, intranodally,intramedullary, intramuscularly, intravenously or intraperitoneally. Insome embodiments, the administering is subcutaneously. In someembodiments, the therapeutically effective amount is at least about 0.1mg/kg, 0.5 mg/kg, 1 mg/kg, 3 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg, 30mg/kg, or 40 mg/kg. In some embodiments, the therapeutically effectiveamount is administered to the subject at least about twice every day oronce every 1-60 days. In some embodiments, the therapeutically effectiveamount is administered to the subject once every 4 weeks. In someembodiments, the autoimmune and/or inflammatory condition is Sjögren'ssyndrome. In some embodiments, the autoimmune and/or inflammatorycondition is rheumatoid arthritis. In some embodiments, the autoimmuneand/or inflammatory condition is IBD. In some embodiments, theautoimmune and/or inflammatory condition is multiple sclerosis. In someembodiments, the multiple sclerosis is clinically isolated syndrome,relapsing-remitting, primary progressive or secondary progressive. Insome embodiments, the autoimmune and/or inflammatory condition isCrohn's disease. In some embodiments, the autoimmune and/or inflammatorycondition is ulcerative colitis. In some embodiments, the autoimmuneand/or inflammatory condition is lupus erythematosus. In someembodiments, the autoimmune and/or inflammatory condition is diabetes,for example type I diabetes. In some embodiments, the IL-7 bindingprotein is used in the manufacture of a medicament for the treatment ofthe autoimmune and/or inflammatory condition. In some embodiments,compounds comprising the IL-7 binding protein are for use in thetreatment of the autoimmune and/or inflammatory condition. In someembodiments, the IL-7 binding protein is used for diagnosis of a diseaseor condition. In some embodiments, the IL-7 binding protein is used fordiagnosis of the autoimmune and/or inflammatory condition. In someembodiments, the IL-7 binding protein is bound to a moiety or anantigenic fragment thereof. In some embodiments, the moiety is IL-7 or afragment thereof. In some embodiments, the IL-7 binding protein isattached directly or indirectly to a solid support. In some embodiments,the IL-7 binding protein is comprised in a kit with instruction for use.In some embodiments, the solid support is comprised in a kit.

Disclosed herein are IL-7 binding proteins or an antigen bindingfragment thereof comprising a heavy chain CDR1 having at least 80%identity to the amino acid sequence set out in SEQ ID NO:6, a heavychain CDR2 having at least 80% identity to the amino acid sequence setout in SEQ ID NO:7 and a heavy chain CDR3 having at least 80% identityto the amino acid sequence set out in SEQ ID NO:8. In some embodiments,the IL-7 binding protein comprises a variable region light chain havingat least 80% identity to the amino acid sequence set out in SEQ ID NO:5In some embodiments, the IL-7 binding protein comprises a variableregion light chain having the amino acid sequence set out in SEQ IDNO:5. In some embodiments, the IL-7 binding protein comprises a heavychain CDR1 comprising the amino acid sequence set out in SEQ ID NO:6, aheavy chain CDR2 comprising the amino acid sequence set out in SEQ IDNO:7 and a heavy chain CDR3 comprising the amino acid sequence set outin SEQ ID NO:8. In some embodiments, the IL-7 binding protein comprisesa light chain CDR1 comprising the amino acid sequence set out in SEQ IDNO:9, a light chain CDR2 comprising the amino acid sequence set out inSEQ ID NO:10 and a light chain CDR3 the amino acid sequence set out inSEQ ID NO:11. In some embodiments, the IL-7 binding protein comprises avariable region heavy chain having at least 80% identity to the aminoacid sequence set out in SEQ ID NO:4. In some embodiments, the IL-7binding protein comprises a variable region heavy chain comprising theamino acid sequence set out in SEQ ID NO:4. In some embodiments, theIL-7 binding protein comprises a light chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein comprises a light chain comprisingthe amino acid sequence set out in SEQ ID NO:3. In some embodiments, theIL-7 binding protein comprises a heavy chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:2. In someembodiments, the IL-7 binding protein comprises a heavy chain comprisingthe amino acid sequence set out in SEQ ID NO:2. In some embodiments, theIL-7 binding protein comprises a heavy chain comprising the amino acidsequence set out in SEQ ID NO:2 and a light chain comprising the aminoacid sequence set out in SEQ ID NO:3. In some embodiments, the IL-7binding protein is an antibody or an antigen-binding portion thereof. Insome embodiments, the antibody is a monoclonal antibody. In someembodiments, the monoclonal antibody is an IgG antibody. In someembodiments, the antibody is a IgG1 isotype antibody. In someembodiments, the monoclonal antibody is an IgG4 isotype antibody. Insome embodiments, the IL-7 binding protein comprises a constant regionsuch that the IL-7 binding protein has reduced ADCC and/or complementactivation or effector functionality. In some embodiments, the IL-7binding protein comprises a heavy chain Fc domain having an alanineresidue at position 235 and/or position 237 according to EU numbering.In some embodiments, the IL-7 binding protein is human, humanized orchimeric. In some embodiments, the IL-7 binding protein is humanized. Insome embodiments, the IL-7 binding protein is human. In someembodiments, the IL-7 binding protein binds to and neutralizes IL-7. Insome embodiments, the IL-7 binding protein binds to native IL-7. In someembodiments, the IL-7 binding protein binds to circulating IL-7. In someembodiments, the IL-7 binding protein is an isolated IL-7 bindingprotein. In some embodiments, the IL-7 binding protein binds to IL-7 andinhibits IL-7 depending IFN-γ or IL-10 secretion from peripheral bloodmononuclear cells with an IC₅₀ of 1 nM or less. In some embodiments, theIL-7 binding protein binds to IL-7 and inhibits IL-7 depending STAT5phosphorylation in CD4+ T cells with an IC₅₀ of 1 nM or less. In someembodiments, the IL-7 binding protein inhibits signaling, activation,cytokine production and proliferation of CD4⁺ T cells and/or CD8⁺ Tcells. In some embodiments, the IL-7 binding protein is a reversibledimer. In some embodiments, the IL-7 binding protein is a dimer. In someembodiments, the IL-7 binding protein is a monomer. In some embodiments,the IL-7 binding protein is for use in a therapy. Further disclosed arenucleic acids encoding the IL-7 binding protein. In some embodiments,the nucleic acid comprises a sequence having at least 80% identity tothe nucleic acid sequence set out in SEQ ID NO:13 encoding the lightchain. In some embodiments, the nucleic acid comprises the nucleic acidsequence set out in SEQ ID NO:13 encoding the light chain. In someembodiments, the nucleic acid comprises a sequence having at least 80%identity to the nucleic acid sequence set out in SEQ ID NO:14 encodingthe heavy chain. In some embodiments, the nucleic acid comprises thenucleic acid sequence set out in SEQ ID NO:14 encoding the heavy chain.In some embodiments, the nucleic acid further comprises a sequencehaving at least 80% identity to the nucleic acid sequence set out in SEQID NO:15 encoding a signal peptide. In some embodiments, disclosedherein are vectors comprising a nucleic acid. In some embodiments, thevector comprises a promoter functional in a mammalian cell. Alsodisclosed are host cells comprising the nucleic acid. In someembodiments, the host cell is a CHO cell. Further disclosed herein aremethods for making the IL-7 binding protein comprising maintaining thehost cell in a medium to produce the IL-7 binding protein and isolatingor purifying the IL-7 binding protein produced by the host cell. In someembodiments, disclosed herein are pharmaceutical compositions comprisingthe IL-7 binding protein. In some embodiment, the pharmaceuticalcompositions comprises a pharmaceutically acceptable carrier orexcipient. In some embodiments, the pharmaceutical composition furthercomprises at least one additional therapeutic agent. In someembodiments, the pharmaceutical composition has a pH of 4.5-7.0. In someembodiments, the pharmaceutical composition has a pH of 5.5, 6.0, 6.2 or6.5. In some embodiments, the pharmaceutical composition or the IL-7binding protein can be used in a method for the treatment of anautoimmune and/or inflammatory condition in a subject in need thereof,the method comprising administering to the subject a therapeuticallyeffective amount of the pharmaceutical composition or the IL-7 bindingprotein. In some embodiments, the administering is transarterially,subcutaneously, intradermally, intratumorally, intranodally,intramedullary, intramuscularly, intravenously or intraperitoneally. Insome embodiments, the administering is subcutaneously. In someembodiments, the therapeutically effective amount is at least about 0.1mg/kg, 0.5 mg/kg, 1 mg/kg, 3 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg, 30mg/kg, or 40 mg/kg. In some embodiments, the therapeutically effectiveamount is administered to the subject at least about twice every day oronce every 1-60 days. In some embodiments, the therapeutically effectiveamount is administered to the subject once every 4 weeks. In someembodiments, the autoimmune and/or inflammatory condition is Sjögren'ssyndrome. In some embodiments, the autoimmune and/or inflammatorycondition is rheumatoid arthritis. In some embodiments, the autoimmuneand/or inflammatory condition is IBD. In some embodiments, theautoimmune and/or inflammatory condition is multiple sclerosis. In someembodiments, the multiple sclerosis is clinically isolated syndrome,relapsing-remitting, primary progressive or secondary progressive. Insome embodiments, the autoimmune and/or inflammatory condition isCrohn's disease. In some embodiments, the autoimmune and/or inflammatorycondition is ulcerative colitis. In some embodiments, the autoimmuneand/or inflammatory condition is lupus erythematosus. In someembodiments, the autoimmune and/or inflammatory condition is diabetes,for example type I diabetes. In some embodiments, the IL-7 bindingprotein is used in the manufacture of a medicament for the treatment ofthe autoimmune and/or inflammatory condition. In some embodiments,compounds comprising the IL-7 binding protein are for use in thetreatment of the autoimmune and/or inflammatory condition. In someembodiments, the IL-7 binding protein is used for diagnosis of a diseaseor condition. In some embodiments, the IL-7 binding protein is used fordiagnosis of the autoimmune and/or inflammatory condition. In someembodiments, the IL-7 binding protein is bound to a moiety or anantigenic fragment thereof. In some embodiments, the moiety is IL-7 or afragment thereof. In some embodiments, the IL-7 binding protein isattached directly or indirectly to a solid support. In some embodiments,the IL-7 binding protein is comprised in a kit with instruction for use.In some embodiments, the solid support is comprised in a kit.

Disclosed herein are IL-7 binding proteins or an antigen bindingfragment thereof comprising a light chain CDR1 having at least 80%identity to the amino acid sequence set out in SEQ ID NO:9, a lightchain CDR2 having at least 80% identity to the amino acid sequence setout in SEQ ID NO:10 and a light chain CDR3 having at least 80% identityto the amino acid sequence set out in SEQ ID NO:11. In some embodiments,the IL-7 binding protein comprises a variable region light chain havingat least 80% identity to the amino acid sequence set out in SEQ ID NO:5.In some embodiments, the IL-7 binding protein comprises a variableregion light chain having the amino acid sequence set out in SEQ IDNO:5. In some embodiments, the IL-7 binding protein comprises a heavychain CDR1 comprising the amino acid sequence set out in SEQ ID NO:6, aheavy chain CDR2 comprising the amino acid sequence set out in SEQ IDNO:7 and a heavy chain CDR3 comprising the amino acid sequence set outin SEQ ID NO:8. In some embodiments, the IL-7 binding protein comprisesa light chain CDR1 comprising the amino acid sequence set out in SEQ IDNO:9, a light chain CDR2 comprising the amino acid sequence set out inSEQ ID NO:10 and a light chain CDR3 the amino acid sequence set out inSEQ ID NO:11. In some embodiments, the IL-7 binding protein comprises avariable region heavy chain having at least 80% identity to the aminoacid sequence set out in SEQ ID NO:4. In some embodiments, the IL-7binding protein comprises a variable region heavy chain comprising theamino acid sequence set out in SEQ ID NO:4. In some embodiments, theIL-7 binding protein comprises a light chain having at least 80%identity to the amino acid set sequence out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein comprises a light chain comprisingthe amino acid sequence set out in SEQ ID NO:3. In some embodiments, theIL-7 binding protein comprises a heavy chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:2. In someembodiments, the IL-7 binding protein comprises a heavy chain comprisingthe amino acid sequence set out in SEQ ID NO:2. In some embodiments, theIL-7 binding protein comprises a heavy chain comprising the amino acidsequence set out in SEQ ID NO:2 and a light chain comprising the aminoacid sequence set out in SEQ ID NO:3. In some embodiments, the IL-7binding protein is an antibody or an antigen-binding portion thereof. Insome embodiments, the antibody is a monoclonal antibody. In someembodiments, the monoclonal antibody is an IgG antibody. In someembodiments, the antibody is a IgG1 isotype antibody. In someembodiments, the monoclonal antibody is an IgG4 isotype antibody. Insome embodiments, the IL-7 binding protein comprises a constant regionsuch that the IL-7 binding protein has reduced ADCC and/or complementactivation or effector functionality. In some embodiments, the IL-7binding protein comprises a heavy chain Fc domain having an alanineresidue at position 235 and/or position 237 according to EU numbering.In some embodiments, the IL-7 binding protein is human, humanized orchimeric. In some embodiments, the IL-7 binding protein is humanized. Insome embodiments, the IL-7 binding protein is human. In someembodiments, the IL-7 binding protein binds to and neutralizes IL-7. Insome embodiments, the IL-7 binding protein binds to native IL-7. In someembodiments, the IL-7 binding protein binds to circulating IL-7. In someembodiments, the IL-7 binding protein is an isolated IL-7 bindingprotein. In some embodiments, the IL-7 binding protein binds to IL-7 andinhibits IL-7 depending IFN-γ or IL-10 secretion from peripheral bloodmononuclear cells with an IC₅₀ of 1 nM or less. In some embodiments, theIL-7 binding protein binds to IL-7 and inhibits IL-7 depending STAT5phosphorylation in CD4+ T cells with an IC₅₀ of 1 nM or less. In someembodiments, the IL-7 binding protein inhibits signaling, activation,cytokine production and proliferation of CD4⁺ T cells and/or CD8⁺ Tcells. In some embodiments, the IL-7 binding protein is a reversibledimer. In some embodiments, the IL-7 binding protein is a dimer. In someembodiments, the IL-7 binding protein is a monomer. In some embodiments,the IL-7 binding protein is for use in a therapy. Further disclosed arenucleic acids encoding the IL-7 binding protein. In some embodiments,the nucleic acid comprises a sequence having at least 80% identity tothe nucleic acid sequence set out in SEQ ID NO:13 encoding the lightchain. In some embodiments, the nucleic acid comprises the nucleic acidsequence set out in SEQ ID NO:13 encoding the light chain. In someembodiments, the nucleic acid comprises a sequence having at least 80%identity to the nucleic acid sequence set out in SEQ ID NO:14 encodingthe heavy chain. In some embodiments, the nucleic acid comprises thenucleic acid sequence set out in SEQ ID NO:14 encoding the heavy chain.In some embodiments, the nucleic acid further comprises a sequencehaving at least 80% identity to the nucleic acid sequence set out in SEQID NO:15 encoding a signal peptide. In some embodiments, disclosedherein are vectors comprising a nucleic acid. In some embodiments, thevector comprises a promoter functional in a mammalian cell. Alsodisclosed are host cells comprising the nucleic acid. In someembodiments, the host cell is a CHO cell. Further disclosed herein aremethods for making the IL-7 binding protein comprising maintaining thehost cell in a medium to produce the IL-7 binding protein and isolatingor purifying the IL-7 binding protein produced by the host cell. In someembodiments, disclosed herein are pharmaceutical compositions comprisingthe IL-7 binding protein. In some embodiment, the pharmaceuticalcompositions comprises a pharmaceutically acceptable carrier orexcipient. In some embodiments, the pharmaceutical composition furthercomprises at least one additional therapeutic agent. In someembodiments, the pharmaceutical composition has a pH of 4.5-7.0. In someembodiments, the pharmaceutical composition has a pH of 5.5, 6.0, 6.2 or6.5. In some embodiments, the pharmaceutical composition or the IL-7binding protein can be used in a method for the treatment of anautoimmune and/or inflammatory condition in a subject in need thereof,the method comprising administering to the subject a therapeuticallyeffective amount of the pharmaceutical composition or the IL-7 bindingprotein. In some embodiments, the administering is transarterially,subcutaneously, intradermally, intratumorally, intranodally,intramedullary, intramuscularly, intravenously or intraperitoneally. Insome embodiments, the administering is subcutaneously. In someembodiments, the therapeutically effective amount is at least about 0.1mg/kg, 0.5 mg/kg, 1 mg/kg, 3 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg, 30mg/kg, or 40 mg/kg. In some embodiments, the therapeutically effectiveamount is administered to the subject at least about twice every day oronce every 1-60 days. In some embodiments, the therapeutically effectiveamount is administered to the subject once every 4 weeks. In someembodiments, the autoimmune and/or inflammatory condition is Sjögren'ssyndrome. In some embodiments, the autoimmune and/or inflammatorycondition is rheumatoid arthritis. In some embodiments, the autoimmuneand/or inflammatory condition is IBD. In some embodiments, theautoimmune and/or inflammatory condition is multiple sclerosis. In someembodiments, the multiple sclerosis is clinically isolated syndrome,relapsing-remitting, primary progressive or secondary progressive. Insome embodiments, the autoimmune and/or inflammatory condition isCrohn's disease. In some embodiments, the autoimmune and/or inflammatorycondition is ulcerative colitis. In some embodiments, the autoimmuneand/or inflammatory condition is lupus erythematosus. In someembodiments, the autoimmune and/or inflammatory condition is diabetes,for example type I diabetes. In some embodiments, the IL-7 bindingprotein is used in the manufacture of a medicament for the treatment ofthe autoimmune and/or inflammatory condition. In some embodiments,compounds comprising the IL-7 binding protein are for use in thetreatment of the autoimmune and/or inflammatory condition. In someembodiments, the IL-7 binding protein is used for diagnosis of a diseaseor condition. In some embodiments, the IL-7 binding protein is used fordiagnosis of the autoimmune and/or inflammatory condition. In someembodiments, the IL-7 binding protein is bound to a moiety or anantigenic fragment thereof. In some embodiments, the moiety is IL-7 or afragment thereof. In some embodiments, the IL-7 binding protein isattached directly or indirectly to a solid support. In some embodiments,the IL-7 binding protein is comprised in a kit with instruction for use.In some embodiments, the solid support is comprised in a kit.

Disclosed herein are pharmaceutical compositions, comprising apharmaceutically-acceptable carrier and an IL-7 binding protein or anantigen binding fragment thereof that exhibits binding specificity forIL-7 at an epitope comprising at least 5 contiguous amino acids of asequence set out in SEQ ID NO:12. In some embodiments, the IL-7 bindingprotein exhibits binding specificity for IL-7 at an epitope comprisingat least 5 contiguous amino acids of a sequence having at least about50%, 60%, 70%, 80%, 90% or 95% identity to the amino acid sequence setout in SEQ ID NO:12. In some embodiments, the IL-7 binding proteinscomprises a light chain CDR1 having at least 80% identity to the aminoacid sequence set out in SEQ ID NO:9, a light chain CDR2 having at least80% identity to the amino acid sequence set out in SEQ ID NO:10 and alight chain CDR3 having at least 80% identity to the amino acid sequenceset out in SEQ ID NO:11. In some embodiments, the IL-7 binding proteincomprises a variable region light chain having at least 80% identity tothe amino acid sequence set out in SEQ ID NO:5. In some embodiments, theIL-7 binding protein comprises a variable region light chain having theamino acid sequence set out in SEQ ID NO:5. In some embodiments, theIL-7 binding protein comprises a heavy chain CDR1 comprising the aminoacid sequence set out in SEQ ID NO:6, a heavy chain CDR2 comprising theamino acid sequence set out in SEQ ID NO:7 and a heavy chain CDR3comprising the amino acid sequence set out in SEQ ID NO:8. In someembodiments, the IL-7 binding protein comprises a light chain CDR1comprising the amino acid sequence set out in SEQ ID NO:9, a light chainCDR2 comprising the amino acid sequence set out in SEQ ID NO:10 and alight chain CDR3 the amino acid sequence set out in SEQ ID NO:11. Insome embodiments, the IL-7 binding protein comprises a variable regionheavy chain having at least 80% identity to the amino acid sequence setout in SEQ ID NO:4. In some embodiments, the IL-7 binding proteincomprises a variable region heavy chain comprising the amino acidsequence set out in SEQ ID NO:4. In some embodiments, the IL-7 bindingprotein comprises a light chain having at least 80% identity to theamino acid sequence set out in SEQ ID NO:3. In some embodiments, theIL-7 binding protein comprises a light chain comprising the amino acidsequence set out in SEQ ID NO:3. In some embodiments, the IL-7 bindingprotein comprises a heavy chain having at least 80% identity to theamino acid sequence set out in SEQ ID NO:2. In some embodiments, theIL-7 binding protein comprises a heavy chain comprising the amino acidsequence set out in SEQ ID NO:2. In some embodiments, the IL-7 bindingprotein comprises a heavy chain comprising the amino acid sequence setout in SEQ ID NO:2 and a light chain comprising the amino acid sequenceset out in SEQ ID NO:3. In some embodiments, the IL-7 binding protein isan antibody or an antigen-binding portion thereof. In some embodiments,the antibody is a monoclonal antibody. In some embodiments, themonoclonal antibody is an IgG antibody. In some embodiments, theantibody is a IgG1 isotype antibody. In some embodiments, the monoclonalantibody is an IgG4 isotype antibody. In some embodiments, the IL-7binding protein comprises a constant region such that the IL-7 bindingprotein has reduced ADCC and/or complement activation or effectorfunctionality. In some embodiments, the IL-7 binding protein comprises aheavy chain Fc domain having an alanine residue at position 235 and/orposition 237 according to EU numbering. In some embodiments, the IL-7binding protein is human, humanized or chimeric. In some embodiments,the IL-7 binding protein is humanized. In some embodiments, the IL-7binding protein is human. In some embodiments, the IL-7 binding proteinbinds to and neutralizes IL-7. In some embodiments, the IL-7 bindingprotein binds to native IL-7. In some embodiments, the pharmaceuticalcomposition further comprises at least one additional therapeutic agent.In some embodiments, the pharmaceutical composition has a pH of 4.5-7.0.In some embodiments, the pharmaceutical composition has a pH of 5.5,6.0, 6.2 or 6.5. In some embodiments, the pharmaceutical composition canbe used in a method for the treatment of an autoimmune and/orinflammatory condition in a subject in need thereof, the methodcomprising administering to the subject a therapeutically effectiveamount of the pharmaceutical composition. In some embodiments, theadministering is transarterially, subcutaneously, intradermally,intratumorally, intranodally, intramedullary, intramuscularly,intravenously or intraperitoneally. In some embodiments, theadministering is subcutaneously. In some embodiments, thetherapeutically effective amount is at least about 0.1 mg/kg, 0.5 mg/kg,1 mg/kg, 3 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg, 30 mg/kg, or 40 mg/kg. Insome embodiments, the therapeutically effective amount is administeredto the subject at least about twice every day or once every 1-60 days.In some embodiments, the therapeutically effective amount isadministered to the subject once every 4 weeks. In some embodiments, theautoimmune and/or inflammatory condition is Sjögren's syndrome. In someembodiments, the autoimmune and/or inflammatory condition is rheumatoidarthritis. In some embodiments, the autoimmune and/or inflammatorycondition is IBD. In some embodiments, the autoimmune and/orinflammatory condition is multiple sclerosis. In some embodiments, themultiple sclerosis is clinically isolated syndrome, relapsing-remitting,primary progressive or secondary progressive. In some embodiments, theautoimmune and/or inflammatory condition is Crohn's disease. In someembodiments, the autoimmune and/or inflammatory condition is ulcerativecolitis. In some embodiments, the autoimmune and/or inflammatorycondition is lupus erythematosus. In some embodiments, the autoimmuneand/or inflammatory condition is diabetes, for example type I diabetes.In some embodiments, the pharmaceutical composition is comprised in akit with instruction for use. In some embodiments, the pharmaceuticalcomposition is a stable liquid aqueous pharmaceutical formulationcomprising an anti-human IL-7 binding protein described herein at aconcentration of 20 to 150 mg/ml, a tonicity agent, a surfactant, and abuffer system having a pH of 4.0 to 8.0.

Disclosed herein are methods for treating an autoimmune and/orinflammatory condition by administering to a subject an IL-7 bindingprotein or an antigen binding fragment thereof. In some embodiments, theIL-7 binding protein comprises a light chain CDR1 having at least 80%identity to the amino acid sequence set out in SEQ ID NO:9, a lightchain CDR2 having at least 80% identity to the amino acid sequence setout in SEQ ID NO:10, and a light chain CDR3 having at least 80% identityto the amino acid sequence set out in SEQ ID NO:11. In some embodiments,the IL-7 binding protein comprises a variable region light chain havingat least 80% identity to the amino acid sequence set out in SEQ ID NO:5.In some embodiments, the IL-7 binding protein comprises a variableregion light chain having the amino acid sequence set out in SEQ IDNO:5. In some embodiments, the IL-7 binding protein comprises a heavychain CDR1 comprising the amino acid sequence set out in SEQ ID NO:6, aheavy chain CDR2 comprising the amino acid sequence set out in SEQ IDNO:7 and a heavy chain CDR3 comprising the amino acid sequence set outin SEQ ID NO:8. In some embodiments, the IL-7 binding protein comprisesa light chain CDR1 comprising the amino acid sequence set out in SEQ IDNO:9, a light chain CDR2 comprising the amino acid sequence set out inSEQ ID NO:10 and a light chain CDR3 the amino acid sequence set out inSEQ ID NO:11. In some embodiments, the IL-7 binding protein comprises avariable region heavy chain having at least 80% identity to the aminoacid sequence set out in SEQ ID NO:4. In some embodiments, the IL-7binding protein comprises a variable region heavy chain comprising theamino acid sequence set out in SEQ ID NO:4. In some embodiments, theIL-7 binding protein comprises a light chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein comprises a light chain comprisingthe amino acid sequence set out in SEQ ID NO:3. In some embodiments, theIL-7 binding protein comprises a heavy chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:2. In someembodiments, the IL-7 binding protein comprises a heavy chain comprisingthe amino acid sequence set out in SEQ ID NO:2. In some embodiments, theIL-7 binding protein comprises a heavy chain comprising the amino acidsequence set out in SEQ ID NO:2 and a light chain comprising the aminoacid sequence set out in SEQ ID NO:3. In some embodiments, the IL-7binding protein is an antibody or an antigen-binding portion thereof. Insome embodiments, the antibody is a monoclonal antibody. In someembodiments, the monoclonal antibody is an IgG antibody. In someembodiments, the antibody is a IgG1 isotype antibody. In someembodiments, the monoclonal antibody is an IgG4 isotype antibody. Insome embodiments, the IL-7 binding protein comprises a constant regionsuch that the IL-7 binding protein has reduced ADCC and/or complementactivation or effector functionality. In some embodiments, the IL-7binding protein comprises a heavy chain Fc domain having an alanineresidue at position 235 and/or position 237 according to EU numbering.In some embodiments, the IL-7 binding protein is human, humanized orchimeric. In some embodiments, the IL-7 binding protein is humanized. Insome embodiments, the IL-7 binding protein is human. In someembodiments, the IL-7 binding protein binds to and neutralizes IL-7. Insome embodiments, the IL-7 binding protein binds to native IL-7. In someembodiments, the administering is transarterially, subcutaneously,intradermally, intratumorally, intranodally, intramedullary,intramuscularly, intravenously or intraperitoneally. In someembodiments, the administering is subcutaneously. In some embodiments,the therapeutically effective amount is at least about 0.1 mg/kg, 0.5mg/kg, 1 mg/kg, 3 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg, 30 mg/kg, or 40mg/kg. In some embodiments, the therapeutically effective amount isadministered to the subject at least about twice every day or once every1-60 days. In some embodiments, the therapeutically effective amount isadministered to the subject once every 4 weeks. In some embodiments, theautoimmune and/or inflammatory condition is Sjögren's syndrome. In someembodiments, the autoimmune and/or inflammatory condition is rheumatoidarthritis. In some embodiments, the autoimmune and/or inflammatorycondition is IBD. In some embodiments, the autoimmune and/orinflammatory condition is multiple sclerosis. In some embodiments, themultiple sclerosis is clinically isolated syndrome, relapsing-remitting,primary progressive or secondary progressive. In some embodiments, theautoimmune and/or inflammatory condition is Crohn's disease. In someembodiments, the autoimmune and/or inflammatory condition is ulcerativecolitis. In some embodiments, the autoimmune and/or inflammatorycondition is lupus erythematosus. In some embodiments, the autoimmuneand/or inflammatory condition is diabetes, for example type I diabetes.

Disclosed herein are use of an IL-7 binding protein or an antigenbinding fragment thereof in the manufacture of a medicament for thetreatment of an autoimmune and/or inflammatory condition. In someembodiments, the IL-7 binding protein comprises a light chain CDR1having at least 80% identity to the amino acid sequence set out in SEQID NO:9, a light chain CDR2 having at least 80% identity to the aminoacid sequence set out in SEQ ID NO:10 and a light chain CDR3 having atleast 80% identity to the amino acid sequence set out in SEQ ID NO:11.In some embodiments, the IL-7 binding protein comprises a variableregion light chain having at least 80% identity to the amino acidsequence set out in SEQ ID NO:5. In some embodiments, the IL-7 bindingprotein comprises a variable region light chain having the amino acidsequence set out in SEQ ID NO:5. In some embodiments, the IL-7 bindingprotein comprises a heavy chain CDR1 comprising the amino acid sequenceset out in SEQ ID NO:6, a heavy chain CDR2 comprising the amino acidsequence set out in SEQ ID NO:7 and a heavy chain CDR3 comprising theamino acid sequence set out in SEQ ID NO:8. In some embodiments, theIL-7 binding protein comprises a light chain CDR1 comprising the aminoacid sequence set out in SEQ ID NO:9, a light chain CDR2 comprising theamino acid sequence set out in SEQ ID NO:10 and a light chain CDR3 theamino acid sequence set out in SEQ ID NO:11. In some embodiments, theIL-7 binding protein comprises a variable region heavy chain having atleast 80% identity to the amino acid sequence set out in SEQ ID NO:4. Insome embodiments, the IL-7 binding protein comprises a variable regionheavy chain comprising the amino acid sequence set out in SEQ ID NO:4.In some embodiments, the IL-7 binding protein comprises a light chainhaving at least 80% identity to the amino acid sequence set out in SEQID NO:3. In some embodiments, the IL-7 binding protein comprises a lightchain comprising the amino acid sequence set out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein comprises a heavy chain having atleast 80% identity to the amino acid sequence set out in SEQ ID NO:2. Insome embodiments, the IL-7 binding protein comprises a heavy chaincomprising the amino acid sequence set out in SEQ ID NO:2. In someembodiments, the IL-7 binding protein comprises a heavy chain comprisingthe amino acid sequence set out in SEQ ID NO:2 and a light chaincomprising the amino acid sequence set out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein is an antibody or anantigen-binding portion thereof. In some embodiments, the antibody is amonoclonal antibody. In some embodiments, the monoclonal antibody is anIgG antibody. In some embodiments, the antibody is a IgG1 isotypeantibody. In some embodiments, the monoclonal antibody is an IgG4isotype antibody. In some embodiments, the IL-7 binding proteincomprises a constant region such that the IL-7 binding protein hasreduced ADCC and/or complement activation or effector functionality. Insome embodiments, the IL-7 binding protein comprises a heavy chain Fcdomain having an alanine residue at position 235 and/or position 237according to EU numbering. In some embodiments, the IL-7 binding proteinis human, humanized or chimeric. In some embodiments, the IL-7 bindingprotein is humanized. In some embodiments, the IL-7 binding protein ishuman. In some embodiments, the IL-7 binding protein binds to andneutralizes IL-7. In some embodiments, the IL-7 binding protein binds tonative IL-7. In some embodiments, the autoimmune and/or inflammatorycondition is Sjögren's syndrome. In some embodiments, the autoimmuneand/or inflammatory condition is rheumatoid arthritis. In someembodiments, the autoimmune and/or inflammatory condition is IBD. Insome embodiments, the autoimmune and/or inflammatory condition ismultiple sclerosis. In some embodiments, the multiple sclerosis isclinically isolated syndrome, relapsing-remitting, primary progressiveor secondary progressive. In some embodiments, the autoimmune and/orinflammatory condition is Crohn's disease. In some embodiments, theautoimmune and/or inflammatory condition is ulcerative colitis. In someembodiments, the autoimmune and/or inflammatory condition is lupuserythematosus. In some embodiments, the autoimmune and/or inflammatorycondition is diabetes, for example type I diabetes.

Disclosed herein are IL-7 binding proteins or an antigen bindingfragment thereof for use in the treatment of an autoimmune and/orinflammatory condition. In some embodiments, the IL-7 binding proteinscomprises a light chain CDR1 having at least 80% identity to the aminoacid sequence set out in SEQ ID NO:9, a light chain CDR2 having at least80% identity to the amino acid sequence set out in SEQ ID NO:10 and alight chain CDR3 having at least 80% identity to the amino acid sequenceset out in SEQ ID NO:11. In some embodiments, the IL-7 binding proteincomprises a variable region light chain having at least 80% identity tothe amino acid sequence set out in SEQ ID NO:5. In some embodiments, theIL-7 binding protein comprises a variable region light chain having theamino acid sequence set out in SEQ ID NO:5. In some embodiments, theIL-7 binding protein comprises a heavy chain CDR1 comprising the aminoacid sequence set out in SEQ ID NO:6, a heavy chain CDR2 comprising theamino acid sequence set out in SEQ ID NO:7 and a heavy chain CDR3comprising the amino acid sequence set out in SEQ ID NO:8. In someembodiments, the IL-7 binding protein comprises a light chain CDR1comprising the amino acid sequence set out in SEQ ID NO:9, a light chainCDR2 comprising the amino acid sequence set out in SEQ ID NO:10 and alight chain CDR3 the amino acid sequence set out in SEQ ID NO:11. Insome embodiments, the IL-7 binding protein comprises a variable regionheavy chain having at least 80% identity to the amino acid sequence setout in SEQ ID NO:4. In some embodiments, the IL-7 binding proteincomprises a variable region heavy chain comprising the amino acidsequence set out in SEQ ID NO:4. In some embodiments, the IL-7 bindingprotein comprises a light chain having at least 80% identity to theamino acid sequence set out in SEQ ID NO:3. In some embodiments, theIL-7 binding protein comprises a light chain comprising the amino acidsequence set out in SEQ ID NO:3. In some embodiments, the IL-7 bindingprotein comprises a heavy chain having at least 80% identity to theamino acid sequence set out in SEQ ID NO:2. In some embodiments, theIL-7 binding protein comprises a heavy chain comprising the amino acidsequence set out in SEQ ID NO:2. In some embodiments, the IL-7 bindingprotein comprises a heavy chain comprising the amino acid sequence setout in SEQ ID NO:2 and a light chain comprising the amino acid sequenceset out in SEQ ID NO:3. In some embodiments, the IL-7 binding protein isan antibody or an antigen-binding portion thereof. In some embodiments,the antibody is a monoclonal antibody. In some embodiments, themonoclonal antibody is an IgG antibody. In some embodiments, theantibody is a IgG1 isotype antibody. In some embodiments, the monoclonalantibody is an IgG4 isotype antibody. In some embodiments, the IL-7binding protein comprises a constant region such that the IL-7 bindingprotein has reduced ADCC and/or complement activation or effectorfunctionality. In some embodiments, the IL-7 binding protein comprises aheavy chain Fc domain having an alanine residue at position 235 and/orposition 237 according to EU numbering. In some embodiments, the IL-7binding protein is human, humanized or chimeric. In some embodiments,the IL-7 binding protein is humanized. In some embodiments, the IL-7binding protein is human. In some embodiments, the IL-7 binding proteinbinds to and neutralizes IL-7. In some embodiments, the IL-7 bindingprotein binds to native IL-7. In some embodiments, the autoimmune and/orinflammatory condition is Sjögren's syndrome. In some embodiments, theautoimmune and/or inflammatory condition is rheumatoid arthritis. Insome embodiments, the autoimmune and/or inflammatory condition is IBD.In some embodiments, the autoimmune and/or inflammatory condition ismultiple sclerosis. In some embodiments, the multiple sclerosis isclinically isolated syndrome, relapsing-remitting, primary progressiveor secondary progressive. In some embodiments, the autoimmune and/orinflammatory condition is Crohn's disease. In some embodiments, theautoimmune and/or inflammatory condition is ulcerative colitis. In someembodiments, the autoimmune and/or inflammatory condition is lupuserythematosus. In some embodiments, the autoimmune and/or inflammatorycondition is diabetes, for example type I diabetes.

Disclosed herein are use of an IL-7 binding protein or an antigenbinding fragment thereof for diagnosis of a disease or condition. Insome embodiments, the IL-7 binding proteins comprises a light chain CDR1having at least 80% identity to the amino acid sequence set out in SEQID NO:9, a light chain CDR2 having at least 80% identity to the aminoacid sequence set out in SEQ ID NO:10 and a light chain CDR3 having atleast 80% identity to the amino acid sequence set out in SEQ ID NO:11.In some embodiments, the IL-7 binding protein comprises a variableregion light chain having at least 80% identity to the amino acidsequence set out in SEQ ID NO:5. In some embodiments, the IL-7 bindingprotein comprises a variable region light chain having the amino acidsequence set out in SEQ ID NO:5. In some embodiments, the IL-7 bindingprotein comprises a heavy chain CDR1 comprising the amino acid sequenceset out in SEQ ID NO:6, a heavy chain CDR2 comprising the amino acidsequence set out in SEQ ID NO:7 and a heavy chain CDR3 comprising theamino acid sequence set out in SEQ ID NO:8. In some embodiments, theIL-7 binding protein comprises a light chain CDR1 comprising the aminoacid sequence set out in SEQ ID NO:9, a light chain CDR2 comprising theamino acid sequence set out in SEQ ID NO:10 and a light chain CDR3 theamino acid sequence set out in SEQ ID NO:11. In some embodiments, theIL-7 binding protein comprises a variable region heavy chain having atleast 80% identity to the amino acid sequence set out in SEQ ID NO:4. Insome embodiments, the IL-7 binding protein comprises a variable regionheavy chain comprising the amino acid sequence set out in SEQ ID NO:4.In some embodiments, the IL-7 binding protein comprises a light chainhaving at least 80% identity to the amino acid sequence set out in SEQID NO:3. In some embodiments, the IL-7 binding protein comprises a lightchain comprising the amino acid sequence set out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein comprises a heavy chain having atleast 80% identity to the amino acid sequence set out in SEQ ID NO:2. Insome embodiments, the IL-7 binding protein comprises a heavy chaincomprising the amino acid sequence set out in SEQ ID NO:2. In someembodiments, the IL-7 binding protein comprises a heavy chain comprisingthe amino acid sequence set out in SEQ ID NO:2 and a light chaincomprising the amino acid sequence set out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein is an antibody or anantigen-binding portion thereof. In some embodiments, the antibody is amonoclonal antibody. In some embodiments, the monoclonal antibody is anIgG antibody. In some embodiments, the antibody is a IgG1 isotypeantibody. In some embodiments, the monoclonal antibody is an IgG4isotype antibody. In some embodiments, the IL-7 binding proteincomprises a constant region such that the IL-7 binding protein hasreduced ADCC and/or complement activation or effector functionality. Insome embodiments, the IL-7 binding protein comprises a heavy chain Fcdomain having an alanine residue at position 235 and/or position 237according to EU numbering. In some embodiments, the IL-7 binding proteinis human, humanized or chimeric. In some embodiments, the IL-7 bindingprotein is humanized. In some embodiments, the IL-7 binding protein ishuman. In some embodiments, the IL-7 binding protein binds to andneutralizes IL-7. In some embodiments, the IL-7 binding protein binds tonative IL-7. In some embodiments, the disease or condition is anautoimmune and/or inflammatory condition. In some embodiments, theautoimmune and/or inflammatory condition is Sjögren's syndrome. In someembodiments, the autoimmune and/or inflammatory condition is rheumatoidarthritis. In some embodiments, the autoimmune and/or inflammatorycondition is IBD. In some embodiments, the autoimmune and/orinflammatory condition is multiple sclerosis. In some embodiments, themultiple sclerosis is clinically isolated syndrome, relapsing-remitting,primary progressive or secondary progressive. In some embodiments, theautoimmune and/or inflammatory condition is Crohn's disease. In someembodiments, the autoimmune and/or inflammatory condition is ulcerativecolitis. In some embodiments, the autoimmune and/or inflammatorycondition is lupus erythematosus. In some embodiments, the autoimmuneand/or inflammatory condition is diabetes, for example type I diabetes.In some embodiments, detection of binding of an IL-7 binding proteindisclosed herein is indicative of a presence of IL-7. In someembodiments, a level of IL-7 is measured in a subject or on/in a sampleobtained from a subject. In some embodiments, measuring a level of IL-7can be performed by a method and assay known in the art. In someembodiments, a level of IL-7 is compared to a reference level of IL-7.In some embodiments, a references level is indicative of a normal,non-diseases, disease, or disease stage. In some embodiments, a level ofIL-7 is measured multiple times in a subject or from multiple samplesobtained from a subject.

Disclosed herein is a solid support comprising an IL-7 binding proteinor an antigen binding fragment thereof. In some embodiments, the solidsupport is an array. In some embodiments, the IL-7 binding protein isattached directly or indirectly to a solid support. In some embodiments,the IL-7 binding proteins comprises a light chain CDR1 having at least80% identity to the amino acid sequence set out in SEQ ID NO:9, a lightchain CDR2 having at least 80% identity to the amino acid sequence setout in SEQ ID NO:10 and a light chain CDR3 having at least 80% identityto the amino acid sequence set out in SEQ ID NO:11. In some embodiments,the IL-7 binding protein comprises a variable region light chain havingat least 80% identity to the amino acid sequence set out in SEQ ID NO:5.In some embodiments, the IL-7 binding protein comprises a variableregion light chain having the amino acid sequence set out in SEQ IDNO:5. In some embodiments, the IL-7 binding protein comprises a heavychain CDR1 comprising the amino acid sequence set out in SEQ ID NO:6, aheavy chain CDR2 comprising the amino acid sequence set out in SEQ IDNO:7 and a heavy chain CDR3 comprising the amino acid sequence set outin SEQ ID NO:8. In some embodiments, the IL-7 binding protein comprisesa light chain CDR1 comprising the amino acid sequence set out in SEQ IDNO:9, a light chain CDR2 comprising the amino acid sequence set out inSEQ ID NO:10 and a light chain CDR3 the amino acid sequence set out inSEQ ID NO:11. In some embodiments, the IL-7 binding protein comprises avariable region heavy chain having at least 80% identity to the aminoacid sequence set out in SEQ ID NO:4. In some embodiments, the IL-7binding protein comprises a variable region heavy chain comprising theamino acid sequence set out in SEQ ID NO:4. In some embodiments, theIL-7 binding protein comprises a light chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein comprises a light chain comprisingthe amino acid sequence set out in SEQ ID NO:3. In some embodiments, theIL-7 binding protein comprises a heavy chain having at least 80%identity to the amino acid sequence set out in SEQ ID NO:2. In someembodiments, the IL-7 binding protein comprises a heavy chain comprisingthe amino acid sequence set out in SEQ ID NO:2. In some embodiments, theIL-7 binding protein comprises a heavy chain comprising the amino acidsequence set out in SEQ ID NO:2 and a light chain comprising the aminoacid sequence set out in SEQ ID NO:3. In some embodiments, the IL-7binding protein is an antibody or an antigen-binding portion thereof. Insome embodiments, the antibody is a monoclonal antibody. In someembodiments, the monoclonal antibody is an IgG antibody. In someembodiments, the antibody is a IgG1 isotype antibody. In someembodiments, the monoclonal antibody is an IgG4 isotype antibody. Insome embodiments, the IL-7 binding protein comprises a constant regionsuch that the IL-7 binding protein has reduced ADCC and/or complementactivation or effector functionality. In some embodiments, the IL-7binding protein comprises a heavy chain Fc domain having an alanineresidue at position 235 and/or position 237 according to EU numbering.In some embodiments, the IL-7 binding protein is human, humanized orchimeric. In some embodiments, the IL-7 binding protein is humanized. Insome embodiments, the IL-7 binding protein is human. In someembodiments, the IL-7 binding protein binds to and neutralizes IL-7. Insome embodiments, the IL-7 binding protein binds to native IL-7. In someembodiments, a device comprises the solid support and a processor fordetecting a signal, the signal is indicative of a binding of a moiety tothe IL-7 binding protein. In some embodiments, the IL-7 binding proteinis comprised in a kit with instruction for use. In some embodiments, thesolid support is comprised in a kit.

Further disclosed herein is a syringe or autoinjector device comprisingan IL-7 binding protein or an antigen binding fragment thereof. In someembodiments, the IL-7 binding proteins comprises a light chain CDR1having at least 80% identity to the amino acid sequence set out in SEQID NO:9, a light chain CDR2 having at least 80% identity to the aminoacid sequence set out in SEQ ID NO:10 and a light chain CDR3 having atleast 80% identity to the amino acid sequence set out in SEQ ID NO:11.In some embodiments, the IL-7 binding protein comprises a variableregion light chain having at least 80% identity to the amino acidsequence set out in SEQ ID NO:5. In some embodiments, the IL-7 bindingprotein comprises a variable region light chain having the amino acidsequence set out in SEQ ID NO:5. In some embodiments, the IL-7 bindingprotein comprises a heavy chain CDR1 comprising the amino acid sequenceset out in SEQ ID NO:6, a heavy chain CDR2 comprising the amino acidsequence set out in SEQ ID NO:7 and a heavy chain CDR3 comprising theamino acid sequence set out in SEQ ID NO:8. In some embodiments, theIL-7 binding protein comprises a light chain CDR1 comprising the aminoacid sequence set out in SEQ ID NO:9, a light chain CDR2 comprising theamino acid sequence set out in SEQ ID NO:10 and a light chain CDR3 theamino acid sequence set out in SEQ ID NO:11. In some embodiments, theIL-7 binding protein comprises a variable region heavy chain having atleast 80% identity to the amino acid sequence set out in SEQ ID NO:4. Insome embodiments, the IL-7 binding protein comprises a variable regionheavy chain comprising the amino acid sequence set out in SEQ ID NO:4.In some embodiments, the IL-7 binding protein comprises a light chainhaving at least 80% identity to the amino acid sequence set out in SEQID NO:3. In some embodiments, the IL-7 binding protein comprises a lightchain comprising the amino acid sequence set out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein comprises a heavy chain having atleast 80% identity to the amino acid sequence set out in SEQ ID NO:2. Insome embodiments, the IL-7 binding protein comprises a heavy chaincomprising the amino acid sequence set out in SEQ ID NO:2. In someembodiments, the IL-7 binding protein comprises a heavy chain comprisingthe amino acid sequence set out in SEQ ID NO:2 and a light chaincomprising the amino acid sequence set out in SEQ ID NO:3. In someembodiments, the IL-7 binding protein is an antibody or anantigen-binding portion thereof. In some embodiments, the antibody is amonoclonal antibody. In some embodiments, the monoclonal antibody is anIgG antibody. In some embodiments, the antibody is a IgG1 isotypeantibody. In some embodiments, the monoclonal antibody is an IgG4isotype antibody. In some embodiments, the IL-7 binding proteincomprises a constant region such that the IL-7 binding protein hasreduced ADCC and/or complement activation or effector functionality. Insome embodiments, the IL-7 binding protein comprises a heavy chain Fcdomain having an alanine residue at position 235 and/or position 237according to EU numbering. In some embodiments, the IL-7 binding proteinis human, humanized or chimeric. In some embodiments, the IL-7 bindingprotein is humanized. In some embodiments, the IL-7 binding protein ishuman. In some embodiments, the IL-7 binding protein binds to andneutralizes IL-7. In some embodiments, the IL-7 binding protein binds tonative IL-7. In some embodiments, the syringe or autoinjector device iscomprised in a kit with instruction for use.

Other aspects and embodiments of the disclosure will be apparent fromthe detailed description that follows.

DESCRIPTION OF DRAWINGS/FIGURES

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIG. 1A: illustrates the concentration of DRSPAI-L7B in animals dosed(i.v.) with a DRSPAI-L7B target dose of 0.1 mg/kg, 1 mg/kg or 10 mg/kg.FIG. 1B and FIG. 1C illustrate the concentration of subcutaneous weeklydoses with a repeat target dose of 30 mg/kg, FIG. 1B following dose 1and FIG. 1C following doses 4. Serum samples were collected at theindicated timepoints and DRSPAI-L7B was quantified by Gyrolabimmunoassay.

FIG. 2A: illustrates % pSTAT5 inhibition of whole blood obtained fromhealthy donors that was stimulated with IL-7 in the presence ofDRSPAI-L7B. FIG. 2B illustrates STAT5 phosphorylation in CD8+ T cells,FIG. 2C CD4+ T cells, FIG. 2D CD3+ T cells, from peripheral bloodmononuclear cells (PBMCs) obtained from healthy donors or IBD patients,stimulated with rhIL-7 in the presence of DRSPAI-L7B or anti-RSVantibody (isotype control).

FIG. 3A: illustrates inhibition of IFN-γ secretion by healthy donorPBMCs treated with increasing concentrations of DRSPAI-L7B in thepresence of rhIL-7 and anti-CD3. FIG. 3B illustrates inhibition of IL-10secretion by healthy donor PBMCs treated with increasing concentrationsof DRSPAI-L7B in the presence of rhIL-7 and anti-CD3. FIG. 3C, FIG. 3D,FIG. 3E, FIG. 3F, FIG. 3G illustrates inhibition of IL-2 by DRSPAI-L7Bin the presence of rhIL-7 and anti-CD3.

FIG. 4A: illustrates IL-17 inhibition in CD4⁺ T_(mem) cells that wereisolated from healthy donor blood, incubated with IL-7 in the presenceof DRSPAI-L7B after being spiked with phorbol myristate acetate(PMA)/ionomycin. FIG. 4B illustrates TNFα inhibition in CD4⁺ T_(mem)cells that were isolated from healthy donor blood, incubated with IL-7in the presence of DRSPAI-L7B after being spiked with PMA/ionomycin.FIG. 4C: illustrates IL-6 inhibition in CD4⁺ T_(mem) cells that wereisolated from healthy donor blood, incubated with IL-7 in the presenceof DRSPAI-L7B after being spiked with PMA/ionomycin.

FIG. 4D: illustrates IL-10 inhibition in CD4⁺ T_(mem) cells that wereisolated from healthy donor blood, incubated with IL-7 in the presenceof DRSPAI-L7B after being spiked with PMA/ionomycin. FIG. 4E:illustrates INFγ inhibition in CD4⁺ T_(mem) cells that were isolatedfrom healthy donor blood, incubated with IL-7 in the presence ofDRSPAI-L7B after being spiked with PMA/ionomycin. FIG. 4F: illustratesCCL3 inhibition in CD4+ T_(mem) cells that were isolated from healthydonor blood, incubated with IL-7 in the presence of DRSPAI-L7B afterbeing spiked with PMA/Ionomycin.

FIG. 5A: illustrates the profile of CD4⁺ lymphocytes from healthycontrols and MS patients profiled by flow cytometry based on CD45RO,CCR7, CD127 and CD25 expression on the cell surface.

FIG. 5B illustrates the profile of CD8⁺ lymphocytes from healthycontrols and MS patients profiled by flow cytometry based on CD45RO,CCR7, CD127 and CD25 expression on the cell surface. FIG. 5C illustratesthe profile of regulatory T cells from healthy controls and MS patientsprofiled by flow cytometry based on CD45RO, CCR7, CD127 and CD25expression on the cell surface.

FIG. 6A: illustrates STAT5 phosphorylation in CD4⁺ T cells from PBMCsobtained from healthy donors or MS patients stimulated with rhIL-7 inthe presence of DRSPAI-L7B or anti-RSV antibody (isotype control). FIG.6B illustrates STAT5 phosphorylation in CD8⁺ T cells from PBMCs obtainedfrom healthy donors or MS patients stimulated with rhIL-7 in thepresence of DRSPAI-L7B or anti-RSV antibody (isotype control).

FIG. 7A: illustrates linear plot and FIG. 7B illustrates logarithmicplot of IL-7 levels in serum samples from healthy controls (HC, n=10),Crohn's disease (n=15), ulcerative colitis (UC, n=15), systemic lupuserythematosus (SLE, n=15) and primary Sjögren's syndrome (pSS, n=15)patients.

DETAILED DESCRIPTION OF THE DISCLOSURE

Inhibition of IL-7 receptor mediated signaling provides a promisingtherapeutic intervention for the treatment of autoimmune and/orinflammatory diseases. In some embodiments, disclosed herein are IL-7binding proteins that reduces B cell activation, decreased autoantibodyproduction and/or decreased B cell antigen presentation. In someembodiments, an IL-7 binding protein disclosed herein binds IL-7,inhibits IL-7 receptor mediated signaling and will not limit developmentor function of regulatory T cells (T_(regs)). In some embodiments, anIL-7 binding protein disclosed herein inhibits signaling, activation,cytokine production and proliferation of both CD4⁺ and CD8⁺ T cells. Insome embodiments, the IL-7 binding protein blockade of IL-7 mediatedsignaling decreases an inflammatory response.

IL-7 Binding Protein

The term “IL-7 mediated signaling”, as used herein, means the biologicaleffect instigated by the IL-7 receptor complex when bound by its ligand,IL-7. IL-7 mediated signaling therefore includes, but is not necessarilylimited to, one or more, or all, of IL-7 induced phosphorylation ofSTAT-5, IL-7 induced expansion of T_(H)17 cells and IL-7 inducedsurvival of T_(H)17 cells.

The term “IL-7 binding protein” as used herein refers to antibodies andother protein constructs, such as domains, which are capable of bindingto IL-7. The terms “IL-7 binding protein” and “anti-IL-7 antigen bindingprotein” are used interchangeably herein. This does not include thenatural cognate receptor.

The term “antibody” is used herein in the broadest sense to refer tomolecules with an immunoglobulin-like domain (for example IgG, IgM, IgA,IgD or IgE) and includes monoclonal, recombinant, polyclonal, chimeric,human, humanized, multispecific antibodies, including bispecificantibodies, and heteroconjugate antibodies; a single variable domain(e.g., a domain antibody (DAB)), antigen binding antibody fragments,Fab, F(ab′)₂, Fv, disulphide linked Fv, single chain Fv,disulphide-linked scFv, diabodies, TANDABS, etc. and modified versionsof any of the foregoing (for a summary of alternative “antibody” formatssee Holliger and Hudson, Nature Biotechnology, 2005, Vol 23, No. 9,1126-1136).

In some embodiments, an IL-7 binding protein disclosed herein may bederived from rat, mouse, primate (e.g. cynomolgus, Old World monkey orGreat Ape) or human. The IL-7 binding protein may be a human, humanizedor chimeric antibody. The IL-7 binding protein may comprise a constantregion, which may be of any isotype or subclass. The constant region maybe of the IgG isotype, for example IgG1, IgG2, IgG3, IgG4 or variantsthereof. The IL-7 binding protein constant region may be IgG1. In someembodiment, the IL-7 binding protein is an IgG1k antibody.

As used herein, “about” means plus or minus 10%.

The term, “full”, “whole” or “intact” antibody, used interchangeablyherein, refers to a heterotetrameric glycoprotein. An intact antibody iscomposed of two identical heavy chains (HCs) and two identical lightchains (LCs) linked by covalent disulphide bonds. This H₂L₂ structurefolds to form three functional domains comprising two antigen-bindingfragments, known as ‘Fab’ fragments, and a ‘Fc’ crystallisable fragment.The Fab fragment is composed of the variable domain at theamino-terminus, variable heavy (VH) or variable light (VL), and theconstant domain at the carboxyl terminus, CH1 (heavy) and CL (light).The Fc fragment is composed of two domains formed by dimerization ofpaired CH2 and CH3 regions. The Fc may elicit effector functions bybinding to receptors on immune cells or by binding C1q, the firstcomponent of the classical complement pathway. The five classes ofantibodies IgM, IgA, IgG, IgE and IgD are defined by distinct heavychain amino acid sequences, which are called μ, α, γ, ε and δrespectively, each heavy chain can pair with either a K or A lightchain. The majority of antibodies in the serum belong to the IgG class,there are four isotypes of human IgG (IgG1, IgG2, IgG3 and IgG4), thesequences of which differ mainly in their hinge region. In someembodiments, an IL-7 binding protein disclosed herein is a human IgG1.In some embodiments, an IL-7 binding protein disclosed herein is adisulfide-linked α2β2 tetramer. In some embodiments, an IL-7 bindingprotein disclosed herein comprises two light (kappa) and two heavy(IgG1) chains.

Fully human antibodies can be obtained using a variety of methods, forexample using yeast-based libraries or transgenic animals (e.g. mice)that can produce repertoires of human antibodies. Yeast presenting humanantibodies on their surface that bind to an antigen of interest can beselected using FACS (Fluorescence-Activated Cell Sorting) based methodsor by capture on beads using labelled antigens. Transgenic animals thathave been modified to express human immunoglobulin genes can beimmunized with an antigen of interest and antigen-specific humanantibodies isolated using B-cell sorting techniques. Human antibodiesproduced using these techniques can then be characterized for desiredproperties such as affinity, developability and selectivity.

In some embodiments, alternative antibody formats can be used.Alternative antibody formats include alternative scaffolds in which theone or more CDRs of the IL-7 antibody can be arranged onto a suitablenon-immunoglobulin protein scaffold or skeleton, such as an affibody, aSpA scaffold, an LDL receptor class A domain, an avimer (see, e.g., U.S.Patent Application Publication Nos. 2005/0053973, 2005/0089932,2005/0164301) or an EGF domain.

The term “domain” refers to a folded polypeptide structure which retainsits tertiary structure independent of the rest of the polypeptide.Generally, domains are responsible for discrete functional properties ofpolypeptides and in many cases may be added, removed or transferred toother polypeptides without loss of function of the remainder of theprotein and/or of the domain.

The term “single variable domain” refers to a folded polypeptide domaincomprising sequences characteristic of antibody variable domains. Ittherefore includes complete antibody variable domains such as VH, VHHand VL and modified antibody variable domains, for example, in which oneor more loops have been replaced by sequences which are notcharacteristic of antibody variable domains, or antibody variabledomains which have been truncated or comprise N- or C-terminalextensions, as well as folded fragments of variable domains which retainat least the binding activity and specificity of the full-length domain.A single variable domain can bind an antigen or epitope independently ofa different variable region or domain. A “domain antibody” or “DAB” maybe considered the same as a “single variable domain”. A single variabledomain may be a human single variable domain, but also includes singlevariable domains from other species such as rodent (for example, asdisclosed in WO 00/29004 A1), nurse shark and Camelid VHH DABs. CamelidVHH are immunoglobulin single variable domain polypeptides that arederived from species including camel, llama, alpaca, dromedary, andguanaco, which produce heavy chain antibodies naturally devoid of lightchains. Such VHH domains may be humanized according to standardtechniques available in the art, and such domains are considered to be“single variable domains”. As used herein, VH includes camelid VHHdomains.

An antigen binding fragment, IL-7 binding protein fragment, functionalfragment, biologically active fragment or an immunologically effectivefragment may comprise partial heavy or light chain variable sequences.Fragments are at least 5, 6, 8 or 10 amino acids in length.Alternatively, the fragments are at least 15, at least 20, at least 50,at least 75, or at least 100 amino acids in length.

An antigen binding fragment may be provided by means of arrangement ofone or more CDRs on non-antibody protein scaffolds. “Protein Scaffold”as used herein includes but is not limited to an immunoglobulin (Ig)scaffold, for example an IgG scaffold, which may be a four chain or twochain antibody, or which may comprise only the Fc region of an antibody,or which may comprise one or more constant regions from an antibody,which constant regions may be of human or primate origin, or which maybe an artificial chimera of human and primate constant regions.

The protein scaffold may be an Ig scaffold, for example an IgG, or IgAscaffold. The IgG scaffold may comprise some or all the domains of anantibody (i.e. CH1, CH2, CH3, VH, VL). The IL-7 binding protein maycomprise an IgG scaffold selected from IgG1, IgG2, IgG3, IgG4 or IgG4PE.For example, the scaffold may be IgG1. The scaffold may consist of, orcomprise, the Fc region of an antibody, or is a part thereof.

The protein scaffold may be a derivative of a scaffold selected from thegroup consisting of CTLA-4, lipocalin, Protein A derived molecules suchas Z-domain of Protein A (Affibody, SpA), A-domain (Avimer/Maxibody);heat shock proteins such as GroEI and GroES; transferrin (trans-body);ankyrin repeat protein (DARPin); peptide aptamer; C-type lectin domain(Tetranectin); human γ-crystallin and human ubiquitin (affilins); PDZdomains; scorpion toxin kunitz type domains of human proteaseinhibitors; and fibronectin/adnectin; which has been subjected toprotein engineering in order to obtain binding to an antigen other thanthe natural ligand.

The term “antagonist antibody” as used herein refers to an antibody orfragment thereof that is capable of fully or partially inhibiting thebiological activity of the antigen, e.g. IL-7, to which it binds forexample by fully or partially blocking binding to a ligand or reducingthe biological activity of the antigen.

“Antigen binding site” refers to a site on an antigen binding proteinwhich is capable of specifically binding to an antigen, this may be asingle variable domain, or it may be paired VH/VL domains as can befound on a standard antibody. Single-chain Fv (ScFv) domains can alsoprovide antigen-binding sites.

The term “chimeric antigen receptor” (“CAR”) as used herein, refers toan engineered receptor which consists of an extracellular antigenbinding domain (which is usually derived from a monoclonal antibody, orfragment thereof, e.g. a VH domain and a VL domain in the form of ascFv), optionally a spacer region, a transmembrane region, and one ormore intracellular effector domains. CARs have also been referred to aschimeric T cell receptors or chimeric immunoreceptors (CIRs). CARs aregenetically introduced into hematopoietic cells, such as T cells, toredirect T cell specificity for a desired cell-surface antigen,resulting in a CAR-T therapeutic.

A “humanized antibody” refers to a type of engineered antibody havingits CDRs derived from a non-human donor immunoglobulin, the remainingimmunoglobulin-derived parts of the molecule being derived from one ormore human immunoglobulin(s). In addition, framework support residuesmay be altered to preserve binding affinity (see, e.g. Queen et al.Proc. Natl Acad Sci USA, 86:10029-10032 (1989), Hodgson et al.Bio/Technology, 9:421 (1991)). A suitable human acceptor antibody may beone selected from a conventional database, e.g. the KABAT database, LosAlamos database, and Swiss Protein database, by homology to thenucleotide and amino acid sequences of the donor antibody. A humanantibody characterized by a homology to the framework regions of thedonor antibody (on an amino acid basis) may be suitable to provide aheavy chain constant region and/or a heavy chain variable frameworkregion for insertion of the donor CDRs. A suitable acceptor antibodycapable of donating light chain constant or variable framework regionsmay be selected in a similar manner. It should be noted that theacceptor antibody heavy and light chains are not required to originatefrom the same acceptor antibody. The prior art describes several ways ofproducing such humanized antibodies—see, for example, EP-A-0239400 andEP-A-054951.

The term “spacer region” as used herein, refers to an oligo- orpolypeptide that functions to link the transmembrane domain to thetarget binding domain. This region may also be referred to as a “hingeregion” or “stalk region”. The size of the spacer can be varieddepending on the position of the target epitope in order to maintain aset distance (e.g. 14 nm) upon CAR:target binding.

The term “transmembrane domain” as used herein refers to the part of theCAR molecule which traverses the cell membrane.

The term “intracellular effector domain” (also referred to as the“signaling domain”) as used herein refers to the domain in the CAR whichis responsible for intracellular signaling following the binding of theantigen binding domain to the target. The intracellular effector domainis responsible for the activation of at least one of the normal effectorfunctions of the immune cell in which the CAR is expressed. For example,the effector function of a T cell can be a cytolytic activity or helperactivity including the secretion of cytokines.

It will be appreciated by a person skilled in the art that VH and/or VLdomains disclosed herein may be incorporated, e.g. in the form of ascFv, into CAR-T therapeutics.

In some embodiments, IL-7 binding proteins of the present disclosureshow cross-reactivity between human IL-7 and IL-7 from another species,such as cynomolgus macaque IL-7. In an embodiment, the IL-7 bindingproteins of the invention specifically bind human and macaque IL-7. Thisis particularly useful, since drug development typically requirestesting of lead drug candidates in mouse systems before the drug istested in humans. The provision of a drug that can bind human andmacaque species allows one to test results in these systems and makeside-by-side comparisons of data using the same drug. This avoids thecomplication of needing to find a drug that works against a macaque IL-7and a separate drug that works against human IL-7, and also avoids theneed to compare results in humans and macaque using non-identical drugs.Cross reactivity between other species used in disease models such asdog or mice is also envisaged.

Optionally, the binding affinity of the IL-7 binding protein for atleast cynomolgus macaque IL-7 and the binding affinity for human IL-7differ by no more than a factor of 2 or 5. In some embodiments, an IL-7binding protein disclosed herein is species specific.

Affinity, also referred to as “binding affinity”, is the strength ofbinding at a single interaction site, i.e. of one molecule, e.g. an IL-7binding protein of the disclosure, to another molecule, e.g. its targetantigen, at a single binding site. The binding affinity of an IL-7binding protein to its target may be determined by equilibrium methods(e.g. enzyme-linked immunoabsorbent assay (ELISA) or radioimmunoassay(RIA)), or kinetics (e.g. surface plasmon resonance analysis using aBIACORE instrument). For example, the BIACORE method described inExample 2 may be used to measure binding affinity.

Avidity, also referred to as functional affinity, is the cumulativestrength of binding at multiple interaction sites, e.g. the sum total ofthe strength of binding of two molecules (or more, e.g. in the case of abispecific or multispecific molecule) to one another at multiple sites,e.g. taking into account the valency of the interaction.

In an embodiment, the equilibrium dissociation constant (KD) of the IL-7binding protein—IL-7 interaction is about 100 nM or less, about 10 nM orless, about 2 nM or less or about 1 nM or less. Alternatively, the KDmay be between about 5 and about 10 nM; or between about 1 and about 2nM. The KD may be between about 1 pM and about 500 pM; or between about500 pM and about 1 nM. In an embodiment, the equilibrium dissociationconstant (KD) of the IL-7 binding protein—IL-7 interaction is 100 nM orless, 10 nM or less, 2 nM or less or 1 nM or less. Alternatively, the KDmay be between 5 and 10 nM; or between 1 and 2 nM. The KD may be between1 pM and 500 pM; or between 500 pM and 1 nM. A skilled person willappreciate that the smaller the KD numerical value, the stronger thebinding. The reciprocal of KD (i.e. 1/KD) is the equilibrium associationconstant (KA) having units M⁻¹. A skilled person will appreciate thatthe larger the KA numerical value, the stronger the binding. In someembodiments, the KD of the IL-7 binding protein disclosed herein isabout 30 to 90 pM. In some embodiments, the KD of the IL-7 bindingprotein disclosed herein is about 30 to about 80 pM, about 30 to about70 pM, about 30 to about 60 pM, about 30 to about 50 pM, about 30 toabout 55 pM or about 30 to about 40 pM. In some embodiments, the KD ofthe IL-7 binding protein disclosed herein is about 40 to about 80 pM,about 40 to about 70 pM, about 40 to about 60 pM, about 40 to about 50pM, about 40 to about 55 pM. In some embodiments, the KD of the IL-7binding protein disclosed herein is about 30 to about 55 pM. In someembodiments, the KD of the IL-7 binding protein disclosed herein isabout 31, about 34, about 46, about 53, about 69, about 73, about 75 orabout 87 pM. In some embodiments, the KD of the IL-7 binding proteindisclosed herein is about 31, about 34, about 46, about 53, about 69,about 73, about 75 or about 87 pM. In some embodiments, the KD of theIL-7 binding protein disclosed herein is about 34 pM. In someembodiments, the KD of the IL-7 binding protein disclosed herein isabout 67 pM. In some embodiments, the KD of the IL-7 binding proteindisclosed herein is about 30 to about 55 pM at 25° C. In someembodiments, the KD of the IL-7 binding protein disclosed herein isabout 36 pM at about 25° C. In some embodiments, the KD of the IL-7binding protein disclosed herein is about 45 to about 90 pM. In someembodiments, the KD of the IL-7 binding protein disclosed herein isabout 45 to about 90 pM at 37° C. In some embodiments, the KD of theIL-7 binding protein disclosed herein is about 67 pM at 37° C. In someembodiments, the KD of the IL-7 binding protein disclosed herein is 30to 90 pM. In some embodiments, the KD of the IL-7 binding proteindisclosed herein is 30 to 80 pM, 30 to 70 pM, 30 to 60 pM, 30 to 50 pM,30 to 55 pM or 30 to 40 pM. In some embodiments, the KD of the IL-7binding protein disclosed herein is 40 to 80 pM, 40 to 70 pM, 40 to 60pM, 40 to 50 pM, 40 to 55 pM. In some embodiments, the KD of the IL-7binding protein disclosed herein is 30 to 55 pM. In some embodiments,the KD of the IL-7 binding protein disclosed herein is 31, 34, 46, 53,69, 73, 75 or 87 pM. In some embodiments, the KD of the IL-7 bindingprotein disclosed herein is 31, 34, 46, 69, 75 or 87 pM. In someembodiments, the KD of the IL-7 binding protein disclosed herein is 34pM. In some embodiments, the KD of the IL-7 binding protein disclosedherein is 67 pM. In some embodiments, the KD of the IL-7 binding proteindisclosed herein is 30 to 55 pM at 25° C. In some embodiments, the KD ofthe IL-7 binding protein disclosed herein is 36 pM at 25° C. In someembodiments, the KD of the IL-7 binding protein disclosed herein is 45to 90 pM. In some embodiments, the KD of the IL-7 binding proteindisclosed herein is 45 to 90 pM at 37° C. In some embodiments, the KD ofthe IL-7 binding protein disclosed herein is 67 pM at 37° C.

In some embodiments, the KD of the IL-7 binding protein disclosed hereinis 31, 34, 46, 53, 69, 73, 75 or 87 pM, plus or minus 15%. In someembodiments, the KD of the IL-7 binding protein disclosed herein is 31,34, 46, 69, 75 or 87 pM, plus or minus 15%. In some embodiments, the KDof the IL-7 binding protein disclosed herein is 34 pM plus or minus 15%.In some embodiments, the KD of the IL-7 binding protein disclosed hereinis 67 pM plus or minus 15%. In some embodiments, the KD of the IL-7binding protein disclosed herein is 30 to 55 pM at 25° C. In someembodiments, the KD of the IL-7 binding protein disclosed herein is 36pM, plus or minus 15%, at 25° C. In some embodiments, the KD of the IL-7binding protein disclosed herein is 45 to 90 pM. In some embodiments,the KD of the IL-7 binding protein disclosed herein is 45 to 90 pM at37° C. In some embodiments, the KD of the IL-7 binding protein disclosedherein is 67 pM plus or minus 15% at 37° C.

A skilled person will appreciate that surface plasmon resonance (SPR) isa suitable method to measure binging affinity and also determine bindingkinetics, see, e.g. Day et al., Direct comparison of bindingequilibrium, thermodynamic, and rate constants determined by surface-and solution-based biophysical methods, Protein Science (2002),11:1017-1025; and, Hearty et al, “Measuring antibody-antigen bindingkinetics using surface plasmon resonance” Methods Mol Biol (2012)907:411-4.

The dissociation rate constant (kd) or “off-rate” describes thestability of the IL-7 binding protein IL-7 complex, i.e. the fraction ofcomplexes that decay per second. For example, a kd of 0.01 s⁻¹ equatesto 1% of the complexes decaying per second. In an embodiment, thedissociation rate constant (kd) is about 1×10⁻³ s⁻¹ or less, about1×10⁻⁴ s⁻¹ or less, about 1×10⁻⁵ s⁻¹ or less, or about 1×10⁻⁶ s⁻¹ orless. The kd may be between about 1×10⁻⁵ s⁻¹ and about 1×10⁻⁴ s⁻¹; orbetween about 1×10⁻⁴ s⁻¹ and about 1×10⁻³ s⁻¹. In some embodiments, thekd of an IL-7 binding protein disclosed herein is about 2.06×10⁻⁴ s⁻¹ orless, about 1.58×10⁻⁴ s⁻¹ or less, about 1.7×10⁻⁴ s⁻¹ or less, or about5.68×10⁻⁴ s⁻¹ or less, about 6.78×10⁻⁴ s⁻¹ or less, about 8.26×10⁻⁴ s⁻¹or less, about 5.15×10⁻⁴ s⁻¹ or less. In some embodiments, the kd of anIL-7 binding protein disclosed herein is about 1.58×10⁻⁴ s⁻¹ or less. Insome embodiments, the kd of an IL-7 binding protein disclosed herein isabout 5.68×10⁻⁴ s⁻¹ or less. In some embodiments, the kd of an IL-7binding protein disclosed herein is about 2.06×10⁻⁴ s⁻¹ or less, about1.58×10⁻⁴ s⁻¹ or less, or about 1.7×10⁻⁴ s⁻¹ or less at 25° C. In someembodiments, the kd of an IL-7 binding protein disclosed herein is about5.68×10⁻⁴ s⁻¹ or less, about 6.78×10⁻⁴ s⁻¹ or less, about 8.26×10⁻⁴ s⁻¹or less, or about 5.15×10⁻⁴ s⁻¹ or less at 37° C.

In an embodiment, the dissociation rate constant (kd) is 1×10⁻³ s⁻¹ orless, 1×10⁻⁴ s⁻¹ or less, 1×10⁻⁵ s⁻¹ or less, or 1×10⁻⁶ s⁻¹ or less. Thekd may be between 1×10⁻⁵ s⁻¹ and 1×10⁻⁴ s⁻¹; or between 1×10⁻⁴ s⁻¹ and1×10⁻³ s⁻¹. In some embodiments, the kd of an IL-7 binding proteindisclosed herein is 2.06×10⁻⁴ s⁻¹ or less, 1.58×10⁻⁴ s⁻¹ or less,1.7×10⁻⁴ s⁻¹ or less, or 5.68×10⁻⁴ s⁻¹ or less, 6.78×10⁻⁴ s⁻¹ or less,8.26×10⁻⁴ s⁻¹ or less, or 5.15×10⁻⁴ s⁻¹ or less. In some embodiments,the kd of an IL-7 binding protein disclosed herein is 1.58×10⁻⁴ s⁻¹ orless. In some embodiments, the kd of an IL-7 binding protein disclosedherein is 5.68×10⁻⁴ s⁻¹ or less.

In some embodiments, the kd of an IL-7 binding protein disclosed hereinis 2.06×10⁻⁴ s⁻¹ or less, 1.58×10⁻⁴ s⁻¹ or less, or 1.7×10⁻⁴ s⁻¹ or lessat 25° C. In some embodiments, the kd of an IL-7 binding proteindisclosed herein is 5.68×10⁻⁴ s⁻¹ or less, 6.78×10⁻⁴ s⁻¹ or less,8.26×10⁻⁴ s⁻¹ or less, or 5.15×10⁻⁴ s⁻¹ or less at 37° C.

The association rate constant (ka) or “on-rate” describes the rate ofIL-7 binding protein—IL-7 complex formation. In an embodiment, theassociation rate constant (ka) is about 6.49×10⁶ M⁻¹s⁻¹, about 4.65×10⁶M⁻¹s⁻¹, about 3.17×10⁶ M⁻¹s⁻¹, about 8.28×10⁶ M⁻¹s⁻¹, about 1.47×10⁷M⁻¹s⁻¹, about 1.10×10⁷ M⁻¹s⁻¹, or about 5.90×10⁶ M⁻¹s⁻¹. In anembodiment, the association rate constant (ka) is about 6.49×10⁶ M⁻¹s⁻¹,4.65×10⁶ M⁻¹s⁻¹ or about 3.17×10⁶ M⁻¹s⁻¹ at 25° C. In an embodiment, theassociation rate constant (ka) is about 8.28×10⁶ M⁻¹s⁻¹, about 1.47×10⁷M⁻¹s⁻¹, about 1.10×10⁷ M⁻¹s⁻¹, or about 5.90×10⁶ M⁻¹s⁻¹ at 37° C. In anembodiment, the association rate constant (ka) is 6.49×10⁶ M⁻¹s⁻¹,4.65×10⁶ M⁻¹s⁻¹, 3.17×10⁶ M⁻¹s⁻¹, 8.28×10⁶ M⁻¹s⁻¹, 1.47×10⁷ M⁻¹s⁻¹,1.10×10⁷ M⁻¹s⁻¹, or 5.90×10⁶ M⁻¹s⁻¹. In an embodiment, the associationrate constant (ka) is 6.49×10⁶ M⁻¹s⁻¹, 4.65×10⁶ M⁻¹s⁻¹ or 3.17×10⁶M⁻¹s⁻¹ at 25° C. In an embodiment, the association rate constant (ka) is8.28×10⁶ M⁻¹ s⁻¹, 1.47×10⁷ M⁻¹s⁻¹, 1.10×10⁷ M⁻¹s⁻¹, or 5.90×10⁶ M⁻¹s⁻¹at 37° C.

The term “neutralizes” as used throughout the present specificationmeans that the biological activity of IL-7 is reduced in the presence ofan IL-7 binding protein as described herein in comparison to theactivity of IL-7 in the absence of the IL-7 binding protein, in vitro orin vivo. Neutralization may be due to one or more of blocking IL-7binding to its receptor, preventing IL-7 from activating its receptor,down regulating IL-7 or its receptor, or affecting effectorfunctionality.

The reduction or inhibition in biological activity may be partial ortotal. A neutralizing IL-7 binding protein may neutralize the activityof IL-7 by lowering the threshold for B cell activation by at least 20%,30% 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 82%, 84%, 86%, 88%, 90%,92%, 94%, 95%, 96%, 97%, 98%, 99% or 100% relative to IL-7 activity inthe absence of the IL-7 binding protein. Neutralization may bedetermined or measured using one or more assays known to the skilledperson or as described herein. For example, FIGS. 2B, 2C, and 2D.

It will be apparent to those skilled in the art that the term “derived”is intended to define not only the source in the sense of it being thephysical origin for the material but also to define material which isstructurally identical to the material but which does not originate fromthe reference source.

By “isolated” it is intended that the molecule, such as an IL-7 bindingprotein, is removed from the environment in which it may be found innature. For example, the molecule may be purified away from substanceswith which it would normally exist in nature. For example, the IL-7binding protein can be purified to at least 95%, 96%, 97%, 98% or 99%,or greater with respect to a culture media containing the IL-7 bindingprotein. The IL-7 binding proteins and antibodies disclosed herein maybe isolated IL-7 binding proteins and antibodies.

“CDRs” are defined as the complementarity determining region amino acidsequences of an antigen binding protein. These are the hypervariableregions of immunoglobulin heavy and light chains. There are three heavychain and three light chain CDRs (or CDR regions) in the variableportion of an immunoglobulin. Thus, “CDRs” as used herein refers to allthree heavy chain CDRs, all three light chain CDRs, all heavy and lightchain CDRs, or at least two CDRs.

Throughout this specification, amino acid residues in variable domainsequences and variable domain regions within full-length antigen bindingsequences, e.g. within an antibody heavy chain sequence or antibodylight chain sequence, are numbered according to the Kabat numberingconvention. Similarly, the terms “CDR”, “CDRL1”, “CDRL2”, “CDRL3”,“CDRH1”, “CDRH2”, “CDRH3” used in the Examples follow the Kabatnumbering convention. For further information, see Kabat et al.,Sequences of Proteins of Immunological Interest, 4th Ed., U.S.Department of Health and Human Services, National Institutes of Health(1987).

Variants

It will be apparent to those skilled in the art that there arealternative numbering conventions for amino acid residues in variabledomain sequences and full-length antibody sequences. There are alsoalternative numbering conventions for CDR sequences, for example thoseset out in Chothia et al. (1989) Nature 342: 877-883. The structure andprotein folding of the IL-7 binding protein may mean that other residuesare considered part of the CDR sequence and would be understood to be soby a skilled person.

Other numbering conventions for CDR sequences available to a skilledperson include “AbM” (University of Bath) and “contact” (UniversityCollege London) methods. The minimum overlapping region using at leasttwo of the Kabat, Chothia, AbM and contact methods can be determined toprovide the “minimum binding unit”. The minimum binding unit may be asub-portion of a CDR.

Table 1 below represents one definition using each numbering conventionfor each CDR or binding unit. The Kabat numbering scheme is used inTable 1 to number the variable domain amino acid sequence. It should benoted that some of the CDR definitions may vary depending on theindividual publication used.

TABLE 1 Minimum Kabat Chothia AbM Contact Binding CDR CDR CDR CDR UnitH1 31-35/ 26-32/ 26-35/ 30-35/ 31-32 35A/35B 33/34 35A/35B 35A/35B H250-65 52-56 50-58 47-58 52-56 H3 95-102 95-102 95-102 93-101 95-101 L124-34 24-34 24-34 30-36 30-34 L2 50-56 50-56 50-56 46-55 50-55 L3 89-9789-97 89-97 89-96 89-96

Accordingly, an IL-7 binding protein is provided, which comprises anyone or a combination of the following CDRs: CDRH1 of SEQ ID NO: 6, CDRH2of SEQ ID NO: 7, CDRH3 of SEQ ID NO: 8, CDRL1 of SEQ ID NO: 9, CDRL2 ofSEQ ID NO: 10, CDRL3 of SEQ ID NO: 11. CDRs may be modified by at leastone amino acid substitution, deletion or addition, wherein the variantIL-7 binding protein substantially retains the biologicalcharacteristics of the unmodified protein, such as binding to IL-7.

It will be appreciated that each of CDR H1, H2, H3, L1, L2, L3 may bemodified alone or in combination with any other CDR, in any permutationor combination. In one embodiment, a CDR is modified by thesubstitution, deletion or addition of up to 3 amino acids, for example 1or 2 amino acids, for example 1 amino acid. Typically, the modificationis a substitution, particularly a conservative substitution, for exampleas shown in Table 2 below.

TABLE 2 Side chain Members Hydrophobic Met, Ala, Val, Leu, Ile Neutralhydrophilic Cys, Ser, Thr Acidic Asp, Glu Basic Asn, Gln, His, Lys, ArgResidues that influence chain orientation Gly, Pro Aromatic Trp, Tyr,Phe

For example, in a variant CDR, the flanking residues that comprise theCDR as part of alternative definition(s) e.g. Kabat or Chothia, may besubstituted with a conservative amino acid residue. In some embodiments,IL-7 binding proteins comprising variant CDRs as described above may bereferred to herein as “functional CDR variants”.

Accordingly, in another embodiment, IL-7 binding proteins providedherein are IL-7 binding proteins which binds to IL-7 and comprises CDRH3of SEQ ID NO:8 or a variant CDRH3 thereof. In an embodiment, the IL-7binding protein comprises CDRH1 of SEQ ID NO:6 or a variant CDRH1thereof, CDRH2 of SEQ ID NO:7 or a variant CDRH2 thereof, CDRH3 of SEQID NO:8 or a variant CDRH3 thereof, CDRL1 of SEQ ID NO:9 or a variantCDRL1 thereof, CDRL2 of SEQ ID NO:10 or a variant CDRL2 thereof, andCDRL3 of SEQ ID NO:11 or a variant CDRL3 thereof. In some embodiments,IL-7 binding proteins disclosed herein, including those with one or morevariant CDRs, may bind to IL-7 and may also neutralize IL-7 activity.

Disclosed herein are IL-7 binding proteins which binds to IL-7 andcomprises a heavy chain variable region of SEQ ID NO:4. The IL-7 bindingprotein may comprise a light chain variable region of SEQ ID NO:5. Insome embodiments, the IL-7 binding protein binds to and neutralizesIL-7. In one embodiment, the IL-7 binding protein comprises a heavychain variable region of SEQ ID NO:4 and a light chain variable regionof SEQ ID NO:5.

The CDRs L1, L2, L3, H1, H2 and H3 tend to structurally exhibit one of afinite number of main chain conformations (canonicals). The particularcanonical structure class of a CDR is defined by both the length of theCDR and by the loop packing, determined by residues located at keypositions in both the CDRs and the framework regions (structurallydetermining residues or SDRs). Martin and Thornton (1996; J Mol Biol263:800-815) have generated an automatic method to define the “keyresidue” canonical templates. Cluster analysis is used to define thecanonical classes for sets of CDRs, and canonical templates are thenidentified by analyzing buried hydrophobics, hydrogen-bonding residues,and conserved glycines and prolines. The CDRs of antibody sequences canbe assigned to canonical classes by comparing the sequences to the keyresidue templates and scoring each template using identity or similaritymatrices.

Based on the canonical class of the DRSPAI-L7B antibody, functionalantibody binding could be predicted to be maintained in the presence ofthe following CDR substitutions, where the amino acid before the Kabatnumber is the original amino acid sequence of and the amino acidsequence at the end of the Kabat number is the substituted amino acid:

-   -   CDRL1 canonicals:        -   K24R        -   S27aN, S27aD, S27aT, S27aE, S31,N, S31T, S31K, S31G        -   L27bV        -   D27cL, D27cY, D27cV, D27cI, D27cS, D27cN, D27sF, D27cH,            D27cG, D27cT        -   Y32F, Y32N, Y32A, Y32H, Y32S, Y32R        -   I33 M, I33L, I33V, I33R        -   N34H    -   CDRL2 canonicals:        -   G51A    -   CDRL3 canonicals:        -   Q89S, Q89G, Q89F, Q89L, Q90N, Q90H        -   S91N, S91F, S91G, S91R, S91D, S91H, S91T, S91Y, S91V        -   N92Y, N92 W, N92T, N92S, N92R, N92Q, N92Q, N92H, N92A, N92D        -   V93E, V93N, V93G, V93H, V93T, V93S, V93R, V93A        -   D94Y, D94T, D94V, D94L, D94H, D94N, D941, D94 W, D94P, D94S        -   L96P, L96Y, L96R, L961, L96 W, L96F    -   CDRH1 canonicals:        -   Y32I, Y32H, Y32F, Y32T, Y32N, Y32C, Y32E, Y32D        -   G33Y, G33A, G33 W, G33T, G33L, G33V        -   V34 M, V34I, V34L, V34T, V34 W        -   H35E, H35N, H35Q H35S, H35Y, H35T    -   CDRH2 canonicals:        -   I51L, I51V, I51T, I51S, I51N, I51 M        -   G55D        -   Y59L    -   CDRH3 canonicals:        -   Y102H, Y102V, Y102I, Y102S, Y102D, Y102G

As discussed above, the particular canonical structure class of a CDR isdefined by both the length of the CDR and by the loop packing,determined by residues located at key positions in both the CDRs and theframework regions. Thus, substitutions may also be made in the frameworkresidues of an IL-7 binding protein of the invention, based on thecanonical class, while retaining a functional antibody. Suchsubstitutions may include (using Kabat numbering):

Light chain: I, L or V at position 2; Q, L or E at position 3; M or L atposition 4; I or V at position 48; and/or

Heavy chain: V, I or L at position 2; L or V at position 4; L, I, M or Vat position 20; T, A, V, G or S at position 24; F, Y, T or G at position27; F, L, I, V or S at position 29; W or Y at position 47; I, M, L, or Vat position 48; I, L, F, M or V at position 69; R, K, V or I at position71, A, L, V, Y or F at position 78; L or M at position 80, Y or F atposition 90, R, K, G, S, H, N, T, A and/or L at position 94.

Thus, the IL-7 binding protein may have any of the above substitutionswithin the stated positions. There may be multiple substitutions pervariant CDR, per heavy or light chain variable region, per heavy orlight chain, and per IL-7 binding protein, and therefore any combinationof substitution may be present in the IL-7 binding protein of theinvention, provided that the canonical structure of the CDR ismaintained. For the avoidance of doubt, the above-describedsubstitutions should not be construed as limiting the possible CDRsubstitutions which may be performed whilst still retaining a functionalanti-IL-7 antibody.

The VH or VL (or HC or LC) sequence disclosed herein may be a variantsequence with up to 10 amino acid substitutions, additions or deletions.For example, the variant sequence may have up to 9, 8, 7, 6, 5, 4, 3, 2or 1 amino acid substitution(s), addition(s) or deletion(s). Thesequence variation may exclude one or more or all of the CDRs, forexample the CDRs are the same as the VH or VL (or HC or LC) sequence andthe variation is in the remaining portion of the VH or VL (or HC or LC)sequence, so that the CDR sequences are fixed and intact.

Alternatively, the heavy chain variable region may have 75% or greater,80% or greater, 85% or greater, 90% or greater, 95% or greater, 98% orgreater, 99% or greater or 100% identity to the amino acid sequence setforth in SEQ ID NO:4; and the light chain variable region may have 75%or greater, 80% or greater, 85% or greater, 90% or greater, 95% orgreater, 98% or greater, 99% or greater, or 100% identity to the aminoacid sequence set forth in SEQ ID NO:5.

The heavy chain variable region may be a variant of the amino acidsequence set forth in SEQ ID NO:4 which may contain 30, 25, 20, 15, 10,9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid substitutions, insertions ordeletions. The light chain variable region may be a variant of the aminoacid sequence set forth in SEQ ID NO:5 which may contain 30, 25, 20, 15,10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid substitutions, insertions ordeletions.

In a particular embodiment, the IL-7 binding protein described hereinbinds to IL-7 and comprises a heavy chain and light chain variabledomain combination of (SEQ ID NO:4) and (SEQ ID NO:5), or an IL-7binding protein which has a heavy and light chain variable domainshaving at least about 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%identity to SEQ ID NO:4 and SEQ ID NO:5, respectively. In an embodiment,the IL-7 binding protein binds to and neutralizes IL-7.

Any of the heavy chain variable regions of the IL-7 binding proteindescribed herein may be combined with a suitable constant region. Any ofthe light chain variable regions of the IL-7 binding protein may becombined with a suitable constant region. A constant region disclosedherein can be a human constant region.

The present disclosure also provides a nucleic acid molecule whichencodes the polypeptide sequence(s) of any one of the IL-7 bindingproteins described herein. The nucleic acid molecule may comprise asequence encoding (i) one or more CDRHs, the heavy chain variablesequence, or the full length heavy chain sequence; and (ii) one or moreCDRLs, the light chain variable sequence, or the full length light chainsequence, with (i) and (ii) on the same nucleic acid molecule.Alternatively, the nucleic acid molecule which encodes an IL-7 bindingprotein described herein may comprise sequences encoding (a) one or moreCDRHs, the heavy chain variable sequence, or the full length heavy chainsequence; or (b) one or more CDRLs, the light chain variable sequence,or the full length light chain sequence, with (a) and (b) on separatenucleic acid molecules. In some embodiments, the nucleic acid comprisesa sequence having at least about 80%, 90%, 95%, 96%, 97%, 98%, 99%, or100% identity to the nucleic acid sequence set out in SEQ ID NO:13encoding the light chain. In some embodiments, the nucleic acidcomprises the nucleic acid sequence set out in SEQ ID NO:13 encoding thelight chain. In some embodiments, the nucleic acid comprises a sequencehaving at least about 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%identity to the nucleic acid sequence set out in SEQ ID NO:14 encodingthe heavy chain. In some embodiments, the nucleic acid comprises thenucleic acid sequence set out in SEQ ID NO:14 encoding the heavy chain.In some embodiments, the nucleic acid further comprises a sequencehaving at least about 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%identity to the nucleic acid sequence set out in SEQ ID NO:15 encoding asignal peptide.

IL-7 binding proteins as described above, for example variants with apartial alteration of the sequence by chemical modification and/orinsertion, deletion or substitution of one or more amino acid residues,or those with 75% or greater, 80% or greater, 85% or greater, 90% orgreater, 95% or greater, 98% or greater, or 99% or greater identity toany of the sequences described above, may display a potency for bindingto IL-7, as demonstrated by EC50 or surface plasmon resonance analysis,of within 10 fold, or within 5 fold of the potency demonstrated byDRSPAI-L7B. DRSPAI-L7B is a human IgG1 that potently (Kd 67 pM) inhibitsIL-7 mediated signalling by binding to interleukin 7 (IL-7). DRSPAI-L7Bis a disulfide-linked α2β2 tetramer consisting of two light (kappa) andtwo heavy (IgG1) chains. The heavy chain constant region contains twopoint mutations, L235A and G237A (LAGA), which reduce binding of the mAbto Fcγ receptors and prevent Fc-mediated effector function including CDCand ADCC. DRSPAI-L7B comprises a heavy chain having the amino acids setout in SEQ ID NO:2 and a light chain having the amino acids set out inSEQ ID NO:3.

In some embodiments, an IL-7 binding protein described herein comprisesa heavy chain having at least 80%, 85%, 90% or 95% sequence identity tothe amino acids set out in SEQ ID NO:19, 21, or 23. In some embodiments,an IL-7 binding protein described herein comprises a heavy chain havingthe amino acids set out in SEQ ID NO:19, 21, or 23. In some embodiments,an IL-7 binding protein described herein comprises a light chain havingat least 80%, 85%, 90% or 95% sequence identity to the amino acids setout in SEQ ID NO: 18, 20, or 22. In some embodiments, an IL-7 bindingprotein described herein comprises a light chain having the amino acidsset out in SEQ ID NO: 18, 20, or 22. In some embodiments, an IL-7binding protein disclosed herein comprises a heavy chain having theamino acids set out in SEQ ID NO:19 and a light chain having the aminoacids set out in SEQ ID NO:18. In some embodiments, an IL-7 bindingprotein disclosed herein comprises a heavy chain having the amino acidsset out in SEQ ID NO:21 and a light chain having the amino acids set outin SEQ ID NO:20. In some embodiments, an IL-7 binding protein disclosedherein comprises a heavy chain having the amino acids set out in SEQ IDNO:23 and a light chain having the amino acids set out in SEQ ID NO:22.

In some embodiments, DRSPAI-L7B reduces survival and activation ofpathogenic and memory T cells in the periphery, block establishment andmaintenance of tertiary lymphoid organs (TLO) via its role indevelopment of lymphoid tissue inducer cells (Lti) and ILC3, and doesnot impact regulatory T cell number or function. In some embodiments,DRSPAI-L7B functions peripherally and will reduce T cell trafficking toTLOs and the brain and thus does not require CNS penetration. Withregard to MS, in some embodiments, DRSPAI-L7B will reduce inflammationand reduce chronic autoimmunity leading to reduction in relapse rate anddelay in progression in both relapse remitting MS (RRMS) and primaryprogressive MS (PPMS).

The term “epitope” as used herein refers to that portion of the antigenthat makes contact with a particular binding domain of the IL-7 bindingprotein, also known as the paratope. An epitope may be linear orconformational/discontinuous. A conformational or discontinuous epitopecomprises amino acid residues that are separated by other sequences,i.e. not in a continuous sequence in the antigen's primary sequenceassembled by tertiary folding of the polypeptide chain. Although theresidues may be from different regions of the polypeptide chain, theyare in close proximity in the three dimensional structure of theantigen. In the case of multimeric antigens, a conformational ordiscontinuous epitope may include residues from different peptidechains. Particular residues comprised within an epitope can bedetermined through computer modelling programs or via three-dimensionalstructures obtained through methods known in the art, such as X-raycrystallography. Epitope mapping can be carried out using varioustechniques known to persons skilled in the art as described inpublications such as Methods in Molecular Biology ‘Epitope MappingProtocols’, Mike Schutkowski and Ulrich Reineke (volume 524, 2009) andJohan Rockberg and Johan Nilvebrant (volume 1785, 2018). Exemplarymethods include peptide based approaches such as pepscan whereby aseries of overlapping peptides are screened for binding using techniquessuch as ELISA or by in vitro display of large libraries of peptides orprotein mutants, e.g. on phage. Detailed epitope information can bedetermined by structural techniques including X-ray crystallography,solution nuclear magnetic resonance (NMR) spectroscopy andcryogenic-electron microscopy (cryo-EM). Mutagenesis, such as alaninescanning, is an effective approach whereby loss of binding analysis isused for epitope mapping. Another method is hydrogen/deuterium exchange(HDX) combined with proteolysis and liquid-chromatography massspectrometry (LC-MS) analysis to characterize discontinuous orconformational epitopes.

In one aspect, provided is an IL-7 binding protein which binds to humanIL-7 at one or more amino acid residues within SEQ ID NO:1. In oneembodiment, provided is an IL-7 binding protein which binds to humanIL-7 at one or more amino acid residues within SEQ ID NO:12. In anotherembodiment, provided is an IL-7 binding protein which binds to humanIL-7 at one or more amino acid residues within SEQ ID NO:16.

In another aspect, provided is an IL-7 binding protein which protectsresidues as set forth in SEQ ID NO: 12 of IL-7 from deuterium exchangein HDX-MS analysis. In one embodiment, provided is an IL-7 bindingprotein which protects residues 67 to 81 (SEQ ID NO:1) of IL-7 fromdeuterium exchange in HDX-MS analysis. In some embodiments, the IL-7binding protein binds to a sequence having at least about 50%, 60%, 70%,80%, 90% or 95% identity to the amino acid sequence set out in SEQ IDNO:12. In some embodiments, the IL-7 binding protein binds to a sequencehaving at least about 50%, 60%, 70%, 80%, 90% or 95% identity to theamino acid sequence set out in SEQ ID NO:16. In some embodiments, theIL-7 binding protein binds to IL-7 at a site that sits adjacent to aIL-7Rα and γ-chain interaction sites on the folded IL-7 protein. In someembodiments, the IL-7 binding protein exhibits binding specificity forIL-7 at an epitope comprising at least 5 contiguous amino acids of asequence set out in SEQ ID NO:12. In some embodiments, the IL-7 bindingprotein exhibits binding specificity for IL-7 at an epitope comprisingat least 5 contiguous amino acids of a sequence set out in SEQ ID NO:16.In some embodiments, the IL-7 binding protein exhibits bindingspecificity for IL-7 at an epitope comprising at least 10 contiguousamino acids of a sequence set out in SEQ ID NO:12. In some embodiments,the IL-7 binding protein exhibits binding specificity for IL-7 at anepitope comprising at least 10 contiguous amino acids of a sequence setout in SEQ ID NO:16.

In some embodiments, an affinity of an IL-7 binding protein to anantigen can be determined by a competitive binding assay. In someembodiments, a competitive binding assay is an immunoassay. In someembodiments, a competitive binding assay is for example, ELISA or aradioimmunoassay.

In some embodiments, reduction or inhibition in biological activity maybe partial or total. A neutralizing IL-7 binding protein disclosedherein may neutralize the activity of IL-7 by at least 20%, 30% 40%,50%, 55%, 60%, 65%, 70%, 75%, 80%, 82%, 84%, 86%, 88%, 90%, 92%, 94%,95%, 96%, 97%, 98%, 99% or 100% relative to IL-7 activity in the absenceof the IL-7 binding protein. Neutralization may be determined ormeasured using one or more assays known to the skilled person or asdescribed herein.

In some embodiments, an IL-7 binding protein disclosed herein targetsmembrane bound IL-7 receptor α (CD127), which upon IL-7 binding forms aheterodimeric receptor with the common γ chain (CD132). In someembodiments, an IL-7 binding protein disclosed herein targets solubleIL-7 receptor α (sCD127).

Competition between the IL-7 binding protein of the invention and areference IL-7 binding protein, e.g. a reference antibody or an IL-7receptor, may be determined by one or more techniques known to theskilled person such as ELISA, FMAT, Surface Plasmon Resonance (SPR) orForteBio Octet Bio-Layer Interferometry (BLI). Such techniques may alsobe referred to as epitope binning. There are several possible reasonsfor this competition: the two proteins may bind to the same oroverlapping epitopes, there may be steric inhibition of binding, orbinding of the first protein may induce a conformational change in theantigen that prevents or reduces binding of the second protein.

The reduction or inhibition in biological activity may be partial ortotal. A neutralising antigen binding protein may neutralise theactivity of <target> by at least 20%, 30% 40%, 50%, 55%, 60%, 65%, 70%,75%, 80%, 82%, 84%, 86%, 88%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, 99% or100% relative to <target> activity in the absence of the antigen bindingprotein.

Neutralisation may be determined or measured using one or more assaysknown to the skilled person or as described herein.

Percent Identity

“Percent identity” between a query nucleic acid sequence and a subjectnucleic acid sequence is the “Identities” value, expressed as apercentage, that is calculated using a suitable algorithm or software,such as BLASTN, FASTA, DNASTAR Lasergene, GeneDoc, Bioedit, EMBOSSneedle or EMBOSS infoalign, over the entire length of the query sequenceafter a pair-wise global sequence alignment has been performed using asuitable algorithm or software, such as BLASTN, FASTA, ClustalW, MUSCLE,MAFFT, EMBOSS Needle, T-Coffee, and DNASTAR Lasergene. Importantly, aquery nucleic acid sequence may be described by a nucleic acid sequenceidentified in one or more claims herein.

“Percent identity” between a query amino acid sequence and a subjectamino acid sequence is the “Identities” value, expressed as apercentage, that is calculated using a suitable algorithm or software,such as BLASTP, FASTA, DNASTAR Lasergene, GeneDoc, Bioedit, EMBOSSneedle or EMBOSS infoalign, over the entire length of the query sequenceafter a pair-wise global sequence alignment has been performed using asuitable algorithm/software such as BLASTP, FASTA, ClustalW, MUSCLE,MAFFT, EMBOSS Needle, T-Coffee, and DNASTAR Lasergene. Importantly, aquery amino acid sequence may be described by an amino acid sequenceidentified in one or more claims herein.

The query sequence may be 100% identical to the subject sequence, or itmay include up to a certain integer number of amino acid or nucleotidealterations as compared to the subject sequence such that the % identityis less than 100%. For example, the query sequence is at least 50, 60,70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% identical to the subjectsequence. Such alterations include at least one amino acid deletion,substitution (including conservative and non-conservative substitution),or insertion, and wherein said alterations may occur at the amino- orcarboxy-terminal positions of the query sequence or anywhere betweenthose terminal positions, interspersed either individually among theamino acids or nucleotides in the query sequence or in one or morecontiguous groups within the query sequence.

The % identity may be determined across the entire length of the querysequence, including the CDRs. Alternatively, the % identity may excludeone or more or all of the CDRs, for example all of the CDRs are 100%identical to the subject sequence and the % identity variation is in theremaining portion of the query sequence, e.g. the framework sequence, sothat the CDR sequences are fixed and intact. In some embodiments, avariant sequence substantially retains the biological characteristics ofthe unmodified protein, such as DRSPAI-L7B.

Modifications

The skilled person will appreciate that, upon production of an IL-7binding protein, such as an antibody in a host cell, post-translationalmodifications may occur. For example, this may include the cleavage ofcertain leader sequences, the addition of various sugar moieties invarious glycosylation patterns, non-enzymatic glycation, deamidation,oxidation, disulfide bond scrambling and other cysteine variants such asfree sulfhydryls, racemized disulfides, thioethers and trisulfide bonds,isomerization, C-terminal lysine clipping, and N-terminal glutaminecyclisation. The disclosure encompasses the use of IL-7 binding proteinsthat have been subjected to, or have undergone, one or morepost-translational modifications. Thus an “IL-7 binding protein” or“antibody” of the invention includes an “IL-7 binding protein” or“antibody”, respectively, as defined earlier that has undergone apost-translational modification such as described herein.

Glycation is a post-translational non-enzymatic chemical reactionbetween a reducing sugar, such as glucose, and a free amine group in theprotein, and is typically observed at the epsilon amine of lysine sidechains or at the N-Terminus of the protein. Glycation can occur duringproduction and storage only in the presence of reducing sugars.

Deamidation can occur during production and storage, is an enzymaticreaction primarily converting asparagine (N) to iso-aspartic acid(iso-aspartate) and aspartic acid (aspartate) (D) at approximately 3:1ratio. This deamidation reaction is therefore related to isomerizationof aspartate (D) to iso-aspartate. The deamidation of asparagine and theisomerization of aspartate, both involve the intermediate succinimide.To a much lesser degree, deamidation can occur with glutamine residuesin a similar manner. Deamidation can occur in a CDR, in a Fab (non-CDRregion), or in the Fc region.

Oxidation can occur during production and storage (i.e. in the presenceof oxidizing conditions) and results in a covalent modification of aprotein, induced either directly by reactive oxygen species orindirectly by reaction with secondary by-products of oxidative stress.Oxidation happens primarily with methionine residues, but may occur attryptophan and free cysteine residues. Oxidation can occur in a CDR, ina Fab (non-CDR) region, or in the Fc region.

Disulfide bond scrambling can occur during production and basic storageconditions. Under certain circumstances, disulfide bonds can break orform incorrectly, resulting in unpaired cysteine residues (—SH). Thesefree (unpaired) sulfhydryls (—SH) can promote shuffling.

The formation of a thioether and racemization of a disulphide bond canoccur under basic conditions, in production or storage, through a betaelimination of disulphide bridges back to cysteine residues via adehydroalanine and persulfide intermediate. Subsequent crosslinking ofdehydroalanine and cysteine results in the formation of a thioether bondor the free cysteine residues can reform a disulphide bond with amixture of D- and L-cysteine.

Trisulfides result from insertion of a sulfur atom into a disulphidebond (Cys-S—S—S-Cys) and are formed due to the presence of hydrogensulphide in production cell culture.

N-terminal glutamine (Q) and glutamate (glutamic acid) (E) in the heavychain and/or light chain is likely to form pyroglutamate (pGlu) viacyclization. Most pGlu formation happens in the production bioreactor,but it can be formed non-enzymatically, depending on pH and temperatureof processing and storage conditions. Cyclization of N-terminal Q or Eis commonly observed in natural human antibodies.

C-terminal lysine clipping is an enzymatic reaction catalyzed bycarboxypeptidases, and is commonly observed in recombinant and naturalhuman antibodies. Variants of this process include removal of lysinefrom one or both heavy chains due to cellular enzymes from therecombinant host cell. Upon administration to the human subject/patientis likely to result in the removal of any remaining C-terminal lysines.

The terms “peptide”, “polypeptide” and “protein” each refers to amolecule comprising two or more amino acid residues. A peptide may bemonomeric or polymeric.

In some embodiments, it may be desirable to modify the effector functionof the IL-7 binding protein, for instance, to enhance ADCC or CDC,half-life, etc. The IL-7 binding protein may have a half-life of atleast 6 hours, at least 1 day, at least 2 days, at least 3 days, atleast 4 days, at least 5 days, at least 7 days, or at least 9 days invivo in humans, or in a murine animal model.

Mutational changes to the Fc effector portion of the antibody can beused to change the affinity of the interaction between the FcRn andantibody to modulate antibody turnover. The half-life of the antibodycan be extended in vivo. This could be beneficial to patient populationsas maximal dose amounts and maximal dosing frequencies could be achievedas a result of maintaining in vivo IC₅₀ for longer periods of time. TheFc effector function of the antibody may be removed, in its entirety orin part, since it may not be desirable to kill those cells expressingCD127. This removal may result in an increased safety profile.

In some embodiments, an IL-7 binding protein comprising a constantregion may have reduced ADCC and/or complement activation or effectorfunctionality. The constant domain may comprise a naturally disabledconstant region of IgG2 or IgG4 isotype or a mutated IgG1 constantdomain. In some embodiments, the IL-7 binding proteins of the inventionmay be Fc disabled. Examples of suitable modifications are described inEP0307434. One way to achieve Fc disablement comprises the substitutionsof alanine residues at positions 235 and 237 (EU index numbering) of theheavy chain constant region, i.e. L235A and G237A (commonly referred toas “LAGA” mutations). Another example comprises substitution withalanines at positions 234 and 235 (EU index numbering), i.e. L234A andL235A (commonly referred to as “LALA” mutations). In some embodiments,the Fc effector function of an IL-7 binding protein disclosed herein hasbeen disabled using the LAGA mutation. Alternatively, the IL-7 bindingprotein may be Fc enabled and not comprise the alanine substitutions atpositions 235 and 237.

Additional alterations and mutations to decrease effector functioninclude: (with reference to IgG1 unless otherwise noted): a glycosylatedN297A or N297Q or N297G; L235E; IgG4:F234A/L235A; and chimericIgG2/IgG4. IgG2: H268Q/V309L/A330S/P331S, and IgG2:V234A/G237A/P238S/H268A/V309L/A330S/P331S can reduce FcγR and C1qbinding (Wang et al. 2018 and U.S. Pat. No. 8,961,967).

Other mutations that decrease effector function includeL234F/L235E/P331S; a chimeric antibody created using the CH1 and hingeregion from human IgG2 and the CH2 and CH3 regions from human IgG4;IgG2m4, based on the IgG2 isotype with four key amino acid residuechanges derived from IgG4 (H268Q, V309L, A330S and P331S); IgG2a whichcontains V234A/G237A/P238S/H268A/V309L/A330S/P331S substitutions toeliminate affinity for Fcγ receptors and C1q complement protein; IgG2m4(H268Q/V309L/A330S/P331S, changes to IgG4); IgG4 (S228P/L234A/L235A);hulgG1 L234A/L235A (AA); hulgG4 S228P/L234A/L235A; IgG1σ(L234A/L235A/G237A/P238S/H268A/A330S/P331S); IgG4σ1(S228P/F234A/L235A/G237A/P238S); and IgG4σ2(S228P/F234A/L235A/G236/G237A/P238S, wherein

denotes a deletion) (Tam et al., Antibodies 2017, 6(3)).

In some embodiments, an IL-7 binding protein disclosed herein maycomprise one or more modifications selected from a mutated constantdomain such that the antibody has enhanced effector functions/ADCCand/or complement activation. Examples of suitable modifications aredescribed in Shields et al. J. Biol. Chem (2001) 276:6591-6604, Lazar etal. PNAS (2006) 103:4005-4010 and U.S. Pat. No. 6,737,056, WO2004063351and WO2004029207. The IL-7 binding protein may comprise a constantdomain with an altered glycosylation profile such that the IL-7 bindingprotein has enhanced effector functions/ADCC and/or complementactivation. Examples of suitable methodologies to produce an IL-7binding protein with an altered glycosylation profile are described inWO2003/011878, WO2006/014679 and EP1229125.

Host and Vector

The IL-7 binding proteins may be prepared by any of a number ofconventional techniques. For example, IL-7 binding proteins may bepurified from cells that naturally express them (e.g., an antibody canbe purified from a hybridoma that produces it), or produced inrecombinant expression systems.

A number of different expression systems and purification regimes can beused to generate the IL-7 binding protein of the invention. Generally,host cells are transformed with a recombinant expression vector encodingthe desired antigen binding protein. The expression vector may bemaintained by the host as a separate genetic element or integrated intothe host chromosome depending on the expression system.

In some embodiments, an expression vector that comprises a nucleic acidmolecule is described herein. Also provided is a recombinant host cellcomprising an expression vector as described herein. An IL-7 bindingprotein described herein may be produced in a suitable host cell. A widerange of host cells can be employed, including Prokaryotes (includingGram negative or Gram positive bacteria, for example Escherichia coli,Bacilli sp., Pseudomonas sp., Corynebacterium sp.), Eukaryotes includingyeast (for example Saccharomyces cerevisiae, Pichia pastoris), fungi(for example Aspergillus sp.), or higher Eukaryotes including insectcells and cell lines of mammalian origin. Examples of cell lines includeChinese Hamster Ovary (CHO) cells, PER.C6, HEK293, HeLa or NSO. In someembodiments, a host cell described herein is a CHO cell, NSO myelomacells, COS cells or SP2 cells. The host cell may be a non-human hostcell. The host cell may be a non-embryonic host cell. Human cells may beused, thus enabling modified human glycosylation patterns.Alternatively, other eukaryotic cell lines may be employed. In someembodiments, selection of suitable mammalian host cells and methods fortransformation, culture, amplification, screening and product productionand purification are known in the art. In some embodiments, the hostcell is a strain of yeast. The host cell may be cultured in a culturemedia, for example serum-free culture media. The IL-7 binding proteinmay be secreted by the host cell into the culture media. The IL-7binding protein can be purified to at least 95% or greater (e.g. 98% orgreater) with respect to the culture media containing the IL-7 bindingprotein.

The host cell may be an isolated host cell. The host cell is usually notpart of a multicellular organism (e.g., plant or animal). The host cellmay be a non-human host cell.

Appropriate cloning and expression vectors for use with bacterial,fungal, yeast, and mammalian host cells are known in the art.

A method for the production of the IL-7 binding protein as describedherein may comprise the step of culturing a host cell and recovering theIL-7 binding protein. In one aspect of the invention, there is provideda method of making an IL-7 binding protein the method comprisingmaintaining a host cell in a medium to produce the IL-7 binding proteinand isolating or purifying the IL-7 binding protein produced by the hostcell.

A recombinant transformed, transfected, or transduced host cell maycomprise at least one expression cassette, whereby the expressioncassette comprises a polynucleotide encoding a heavy chain of the IL-7binding protein described herein and further comprises a polynucleotideencoding a light chain of the IL-7 binding protein described herein.Alternatively, a recombinant transformed, transfected or transduced hostcell may comprise at least one expression cassette, whereby a firstexpression cassette comprises a polynucleotide encoding a heavy chain ofthe IL-7 binding protein described herein and further comprise a secondcassette comprising a polynucleotide encoding a light chain of the IL-7binding protein described herein. A stably transformed host cell maycomprise a vector comprising one or more expression cassettes encoding aheavy chain and/or a light chain of the IL-7 binding protein describedherein or fragments thereof. For example, such host cells may comprise afirst vector encoding the light chain and a second vector encoding theheavy chain.

The cells can be cultured under conditions that promote expression ofthe antigen binding protein using a variety of equipment such as shakeflasks, spinner flasks, and bioreactors. The polypeptide is recovered byconventional protein purification procedures. Protein purificationprocedures typically consist of a series of unit operations comprised ofvarious filtration and chromatographic processes developed toselectively concentrate and isolate the antigen binding protein. Thepurified antigen binding protein may be formulated in a pharmaceuticallyacceptable composition.

STATEMENT OF USE

In one aspect, an IL-7 binding protein described herein is for use intherapy. An IL-7 binding protein described herein can be used in thetreatment of diseases or conditions for which an IL-7 inhibitor isindicated, for example inflammatory or autoimmune diseases. In someembodiments, IL-7 inhibition by an IL-7 binding protein described hereinimpacts the survival, expansion and function of autoreactive effector Tcells, while sparing regulatory T lymphocytes. In some embodiments, IL-7inhibition by an IL-7 binding protein described herein inhibits theformation of ectopic lymphoid tissue. In some embodiments, IL-7inhibition by an IL-7 binding protein described herein may help torestore homeostasis by inhibiting innate lymphoid cell (ILC) survival.

In some embodiments, an IL-7 binding protein disclosed herein is capableof antagonizing the biological effect of IL-7 and is capable ofantagonizing at least one of IL-7R-mediated T_(H)17 expansion, andIL-7R-mediated T_(H)17 survival. The term inhibit, antagonize andneutralize are used herein synonymously. No term is intended to suggestthe requirement of total neutralization; partialneutralization—corresponding to a reduction but not complete abolitionof the biological effect—is also contemplated.

At a molecular level, T_(H)17 expansion and/or survival can be observedby an increase in IL-17 production by a population of CD4+ T cells (orby a population of T_(H)17 cells). In an embodiment, therefore, the IL-7binding proteins disclosed herein reduces IL-17 production by apopulation of CD4+ T cells. IL-7 receptor mediated T_(H)17 expansion andsurvival can also be observed by an increase in IFN-γ production by apopulation of CD4+ T cells (or by a population of T_(H)17 cells). Thus,in an embodiment, the IL-7 binding proteins disclosed herein antagonize(reduce) IFN-γ production by a population of CD4+ T cells. At amolecular level, the IL-7 binding proteins disclosed herein may inhibitIL-7 receptor mediated STAT-5 phosphorylation.

In some embodiments, at the molecular level, one can observe and measurethe blocking effect of the IL-7 binding proteins of described herein byassays such as IL-7-induced P-STAT5 or Bcl-2. In some embodiments, atthe cellular level, one can observe and measure the blocking effect byassays such as T_(H)17 secretion of IL-17 or IFNγ. Exemplary assays aredescribed in PCT application number PCT/US2009/053136 (WO2010/017468).In an exemplary pSTAT-5 assay, PBMCs can be stimulated with IL-7 in thepresence and absence of a test agent. Cells can be subsequently assessedquantitatively for the level of pSTAT-5, e.g. by staining for pSTAT-5(e.g. with a labelled anti-pSTAT-5 antibody, such as ALEXA FLUOR 647Mouse Anti-Stat5 (pY694, BD [#612599])) followed by fluorescenceactivated cell sorting. The levels of phosphorylated STAT-5 could alsobe determined by ELISA. Those agents which reduce the level ofphosphorylated STAT-5 may be potential therapeutic candidates forautoimmune disease.

In some embodiments, the disclosure provides a method for the treatmentof an autoimmune disease in a human subject, comprising administering tothe subject an IL-7 binding protein in an amount sufficient to reduceIL-7R-mediated STAT-5 phosphorylation. In some embodiments, an IL-7binding protein disclosed herein blocks or inhibits IL-7 mediatedphosphorylation of STAT5 directly downstream of IL-7R. In someembodiments, an IL-7 binding protein disclosed herein downregulatessurface expression of activation markers and chemokine receptorsresponsible for lymphocyte trafficking to the CNS on active T_(H)1cells.

An antagonist, such as the antigen-binding protein of the disclosure maybe capable of reducing levels of phosphorylated STAT-5 by at least 20%,50%, 75%, 80%, 85%, 90%, 95% or 100% when compared to STAT-5 levels inthe absence of the antagonist, or when compared to a negative control,or untreated cells. The antagonist may have an IC₅₀ of 50 μg/ml, 25μg/ml or less, 10 μg/ml or less, 5 μg/ml or less, or 2 μg/ml or less. Inan embodiment, the antagonist has an IC₅₀ of less than or equal to 1μg/ml, less than or equal to 0.75 μg/ml, less than or equal to 0.5μg/ml, less than or equal to 0.25 μg/ml, or less than or equal to 0.1μg/ml. In one embodiment, the antagonist has an IC₅₀ of less than orequal to 50 ng/ml, less than or equal to 40 ng/ml, less than or equal to30 ng/ml, less than or equal to 20 ng/ml or less than or equal to 10ng/ml. In one embodiment, the antagonist has an IC₅₀ of 5 ng/ml.

An antagonist disclosed herein may be particularly effective ininhibiting the expansion of T_(H)17 cells. Expansion of T_(H)17 cellscan be determined in a T_(H)17 expansion assay, which can comprisestimulating a population of naïve T cells to expand in the presence andabsence of a test agent, followed by stimulating the cells to produceIL-17 and assessing the level of IL-17 produced by the cells in thepresence and absence of the test agent. In an exemplary assay, humanCD4+ T cells can be differentiated into T_(H)17 by stimulation with Tcell receptor activation in the presence of IL-1, IL-6, and IL-23. After5 days of differentiation, CCR6+ cells can be sorted out to produce anenriched T_(H)17 population. This population can then be stimulated withhuman IL-7 and the increase in IL-17 and IFN-γ in the supernatant can bedetermined. The ability of a test agent, such as an antigen bindingfragment of the present disclosure, to inhibit induction of IL-7R byIL-7 can be determined as the presence of an antagonist of thisinteraction during the incubation period should prevent the expansion ofthe T_(H)17 cells leading to the reduction of IL-17 and IFN-γproduction.

The IL-7 binding proteins may be capable of from 20% or more inhibitionof IL-17 secretion in such an assay, versus a negative control. Moretypically, the IL-7 binding protein is capable of from 50%, from 75%,from 85% or from 90% or more inhibition of IL-17 secretion versus thecontrol. The IL-7 binding protein may, in some embodiments, exhibit anIC₅₀ of less than or equal to 50 μg/ml in the assay. In otherembodiments, the IC₅₀ may be less than or equal to 20 μg/ml, 10 μg/ml or5 μg/ml. Thus, in another aspect, the disclosure provides a method forthe treatment of an autoimmune disease or inflammatory disorder,comprising administering to a patient an IL-7 binding protein disclosedherein in an amount sufficient to reduce the T_(H)17 cell count in thepatient.

In some embodiments, an IL-7 binding protein disclosed herein is for thetreatment of a subject. The terms “individual”, “subject” and “patient”are used herein interchangeably. The subject is typically a human. Thesubject may also be a mammal, such as a mouse, rat or primate (e.g. amarmoset or monkey). The subject can be a non-human animal. The IL-7binding proteins may also have veterinary use. The subject to be treatedmay be a farm animal for example, a cow or bull, sheep, pig, ox, goat orhorse or may be a domestic animal such as a dog or cat. The animal maybe any age, or a mature adult animal. In some embodiments, treatment maybe therapeutic, prophylactic or preventative. The subject may be one whois in need thereof. Those in need of treatment may include individualsalready suffering from a medical disease in addition to those who maydevelop the disease in the future.

Thus, the IL-7 binding protein described herein can be used forprophylactic or preventative treatment. In this case, the IL-7 bindingprotein described herein is administered to an individual in order toprevent or delay the onset of one or more aspects or symptoms of adisease. The subject can be asymptomatic. The subject may have a geneticpredisposition to the disease. In some embodiments, a prophylacticallyeffective amount of the IL-7 binding protein is administered to such anindividual. In some embodiments, a prophylactically effective amount isan amount which prevents or delays the onset of one or more aspects orsymptoms of a disease described herein.

The IL-7 binding protein described herein may also be used in methods oftherapy. The term “therapy” encompasses alleviation, reduction, orprevention of at least one aspect or symptom of a disease. For example,the IL-7 binding protein described herein may be used to ameliorate orreduce one or more aspects or symptoms of a disease described herein.

In some embodiments, an IL-7 binding protein described herein is used inan effective amount for therapeutic, prophylactic or preventativetreatment. In some embodiments, a therapeutically effective amount ofthe IL-7 binding protein described herein is an amount effective toameliorate or reduce one or more aspects or symptoms of the disease. Insome embodiments, the IL-7 binding protein described herein may also beused to treat, prevent, or cure the disease described herein. In someembodiments, an IL-7 binding protein described herein have a generallybeneficial effect on the subject's health, for example it can increasethe subject's expected longevity.

The IL-7 binding protein described herein need not affect a completecure or eradicate every symptom or manifestation of the disease toconstitute a viable therapeutic treatment. As is recognized in thepertinent field, drugs employed as therapeutic agents may reduce theseverity of a given disease state but need not abolish everymanifestation of the disease to be regarded as useful therapeuticagents. Similarly, a prophylactically administered treatment need not becompletely effective in preventing the onset of a disease in order toconstitute a viable prophylactic agent. Simply reducing the impact of adisease (for example, by reducing the number or severity of itssymptoms, or by increasing the effectiveness of another treatment, or byproducing another beneficial effect), or reducing the likelihood thatthe disease will occur (for example by delaying the onset of thedisease) or worsen in a subject, is sufficient.

In some embodiments, an IL-7 binding protein described herein may beused in a therapy to treat a subject having or suspected of having adisease or condition described herein. In some embodiments, an IL-7binding protein described herein is administered to a subject having orsuspected of having a disease or condition described herein. In someembodiments, an IL-7 binding protein described herein may be used in thetherapy of multiple sclerosis (MS) and in other autoimmune orinflammatory diseases, particularly those in which pathogenic T_(H)17cells are implicated. In some embodiments, an IL-7 binding proteindescribed herein may be used in the therapy of rheumatoid arthritis,psoriasis, Behcet's disease, diabetes, for example type I diabetes andsystemic lupus erythematosus (SLE).

Inhibition of IL-7-induced IL-7R-mediated signaling may be useful in thetreatment of inflammatory (non-autoimmune) diseases in which elevatedIL-17 or IL-2 has been implicated, such as asthma. Accordingly,inflammatory and/or autoimmune diseases that may be treated by an IL-7binding protein disclosed herein include inflammatory skin diseasesincluding psoriasis and atopic dermatitis; systemic scleroderma andsclerosis; inflammatory bowel disease (IBD); Crohn's disease; ulcerativecolitis; ischemic reperfusion disorders including surgical tissuereperfusion injury, myocardial ischemic conditions such as myocardialinfarction, cardiac arrest, reperfusion after cardiac surgery andconstriction after percutaneous transluminal coronary angioplasty,stroke, and abdominal aortic aneurysms; cerebral edema secondary tostroke; cranial trauma, hypovolemic shock; asphyxia; adult respiratorydistress syndrome; acute-lung injury; Behcet's Disease; dermatomyositis;polymyositis; multiple sclerosis (MS); dermatitis; meningitis;encephalitis; uveitis; osteoarthritis; lupus nephritis; autoimmunediseases such as rheumatoid arthritis (RA), spondyloarthritis, Sjögren'ssyndrome, vasculitis; diseases involving leukocyte diapedesis; centralnervous system (CNS) inflammatory disorder, multiple organ injurysyndrome secondary to septicemia or trauma; alcoholic hepatitis;bacterial pneumonia; antigen-antibody complex mediated diseasesincluding glomerulonephritis; sepsis; sarcoidosis; immunopathologicresponses to tissue/organ transplantation; inflammations of the lung,including pleurisy, alveolitis, vasculitis, pneumonia, chronicbronchitis, bronchiectasis, diffuse panbronchiolitis, hypersensitivitypneumonitis, idiopathic pulmonary fibrosis (IPF), and cystic fibrosis;psoriatic arthritis; neuromyelitis optica, Guillain-Barre syndrome(GBS), COPD, type 1 diabetes, etc.

In particular, an IL-7 binding protein disclosed herein may be useful inthe therapy of multiple sclerosis, in all its forms, includingneuromyelitis optica. In some embodiment, an IL-7 binding proteindisclosed herein is useful in the therapy of multiple sclerosisclassified as clinically-isolated syndrome (CIS); primary-progressive(PPMS); relapsing-remitting (RRMS) and/or secondary progressive (SPMS).In some embodiments, treatment with an IL-7 binding protein disclosedherein is predicted to be most efficacious when administered in thecontext of active inflammatory disease, i.e. when used in the treatmentof clinically isolated syndrome or relapsing forms of MS. These stagesof disease can be defined clinically and/or by imaging criteria such asgadolinium enhancement or other more sensitive techniques, and/or otheras yet undefined biomarkers of active disease. Particularly, an IL-7binding protein disclosed herein can be used to treat RRMS (viaintravenous, subcutaneous, oral or intramuscular delivery) when subjectsare entering or are in relapse. In an embodiment, an IL-7 bindingprotein disclosed herein is administered to a subject at the onset ofrelapse, or within 1 hr, 2 hrs, 3 hrs, 6 hrs, 12 hrs, 24 hrs, 2 days, 3days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days from theonset of relapse.

In some embodiment, an autoimmune and/or inflammatory disease isselected from IBD, rheumatoid arthritis (RA), Sjögren's syndrome,Crohn's disease, diabetes, for example type I diabetes, systemic lupuserythematosus (SLE) and ulcerative colitis. In another embodiment, theautoimmune and/or inflammatory condition is rheumatoid arthritis (RA).In another embodiment, the autoimmune and/or inflammatory condition isSjögren's syndrome. In another embodiment, the autoimmune and/orinflammatory condition is systemic lupus erythematosus (SLE). In anotherembodiment, the autoimmune and/or inflammatory condition is ulcerativecolitis. In another embodiment, the autoimmune and/or inflammatorycondition is diabetes, for example type I diabetes.

In some embodiments, provided herein is an IL-7 binding protein for usein the treatment of a disease or condition disclosed herein. In someembodiments, provided herein are IL-7 binding proteins for use in thetreatment of an autoimmune and/or inflammatory disease. In someembodiments, provided herein are IL-7 binding proteins for use in thetreatment of multiple sclerosis (MS). In some embodiments, disclosedherein is are IL-7 binding protein for use in the treatment of relapsingremitting MS, secondary progressive MS, and/or primary progressive MS.In some embodiments, provided herein is an IL-7 binding protein for usein the treatment of rheumatoid arthritis (RA). In some embodiments,provided herein is an IL-7 binding protein for use in the treatment ofSjögren's syndrome. In some embodiments, provided herein is an IL-7binding protein for use in the treatment of systemic lupus erythematosus(SLE). In some embodiments, provided herein is an IL-7 binding proteinfor use in the treatment of ulcerative colitis. In some embodiments,provided herein is an IL-7 binding protein for use in the treatment ofdiabetes, for example type I diabetes.

In some embodiments, provided herein is the use of an IL-7 bindingprotein in the manufacture of a medicament for the treatment of adisease or condition disclosed herein. In some embodiments, providedherein is the use of an IL-7 binding protein in the manufacture of amedicament for the treatment of an autoimmune and/or inflammatorydisease. In some embodiments, provided herein is the use of an IL-7binding protein in the manufacture of a medicament for the treatment ofmultiple sclerosis. In some embodiments, provided herein is the use ofan IL-7 binding protein in the manufacture of a medicament for thetreatment of rheumatoid arthritis (RA). In some embodiments, providedherein is the use of an IL-7 binding protein in the manufacture of amedicament for the treatment of Sjögren's syndrome. In some embodiments,provided herein is the use of an IL-7 binding protein in the manufactureof a medicament for the treatment of systemic lupus erythematosus (SLE).In some embodiments, provided herein is the use of an IL-7 bindingprotein in the manufacture of a medicament for the treatment ofulcerative colitis. In some embodiments, provided herein is the use ofan IL-7 binding protein in the manufacture of a medicament for thetreatment of diabetes, for example type I diabetes.

Also provided is a method for treatment of an autoimmune and/orinflammatory condition in a subject in need thereof, comprisingadministering to the subject a therapeutically effective amount of anIL-7 binding protein disclosed herein. In some embodiments, provided isa method for treating rheumatoid arthritis (RA) in a subject in needthereof, comprising administering to said subject a therapeuticallyeffective amount of an IL-7 binding protein disclosed herein. In someembodiments, provided is a method for treating Sjögren's syndrome in asubject in need thereof, comprising administering to said subject atherapeutically effective amount of an IL-7 binding protein disclosedherein. In some embodiments, provided is a method for treating systemiclupus erythematosus (SLE) in a subject in need thereof, comprisingadministering to said subject a therapeutically effective amount of anIL-7 binding protein disclosed herein. In some embodiments, provided isa method for treating ulcerative colitis in a subject in need thereof,comprising administering to said subject a therapeutically effectiveamount of an IL-7 binding protein disclosed herein. In some embodiments,provided herein is a method for treating multiple sclerosis in a subjectin need thereof, comprising administering to said subject atherapeutically effective amount of an IL-7 binding protein disclosedherein. In some embodiments, provided herein is a method for treatingdiabetes, for example type I diabetes in a subject in need thereof,comprising administering to said subject a therapeutically effectiveamount of an IL-7 binding protein disclosed herein.

In some embodiments, disclosed herein is a method for treating multiplesclerosis in a patient comprising administering an IL-7 binding proteindisclosed herein to the patient, wherein the patient is suffering fromrelapsing remitting multiple sclerosis. In some embodiments, disclosedherein is a method for treating an autoimmune or inflammatory disease ina human subject, comprising administering to the subject an IL-7 bindingprotein disclosed herein to the patient in an amount effective to reducethe ratio of T_(H)17 cells relative to T_(H)1 cells. In someembodiments, disclosed herein is a method for treating an autoimmune orinflammatory disease in a human subject, comprising administering to thesubject an IL-7 binding protein disclosed herein to the patient in anamount effective to reduce the ratio of T_(H) cells relative to (Foxp3+)T_(reg) cells.

In some embodiments, a treatment may comprise further monitoring of adisease or condition of a subject. A treatment may comprise a singletreatment. A treatment may comprise a recurring treatment. A treatmentmay comprise a recurring treatment over a remaining lifespan of asubject. A treatment may comprise a daily treatment. A treatment maycomprise a biweekly treatment. In some embodiments, a treatment may beselected based on an assessment of a patient or a sample obtained fromthe patient.

Pharmaceutical Compositions/Routes of Administration/Dosages

IL-7 binding protein as described herein may be incorporated intopharmaceutical compositions for use in the treatment of the humandiseases described herein. In one embodiment, the pharmaceuticalcomposition comprises an IL-7 binding protein in combination with one ormore pharmaceutically acceptable carriers and/or excipients.

Such compositions comprise a pharmaceutically acceptable carrier asknown and called for by acceptable pharmaceutical practice. In oneembodiment, the pharmaceutical composition comprises apharmaceutically-acceptable carrier or excipient and an IL-7 bindingprotein that exhibits binding for IL-7 at an epitope comprising at least5 contiguous amino acids of a sequence set out in SEQ ID NO:12 or SEQ IDNO:16

Pharmaceutical compositions of the disclosures may be used fortherapeutic or prophylactic applications. In some embodiments, providedare pharmaceutical compositions comprising an IL-7 binding protein and apharmaceutically acceptable carrier or excipient thereof. In anotherembodiment, provided are pharmaceutical compositions comprising 1-500 mgof an IL-7 binding protein disclosed herein. In another embodiment,provided are pharmaceutical composition comprising 20-300 mg of an IL-7binding protein disclosed herein. In another embodiment, provided arepharmaceutical compositions comprising 50-200 mg of an IL-7 bindingprotein disclosed herein. In a further embodiment, provided herein arepharmaceutical compositions comprising 50-200 mg of a IL-7 bindingprotein which is an antibody comprising a light chain amino acidsequence as set out in SEQ ID NO:3 and a heavy chain amino acid sequenceas set out in SEQ ID NO:2. In a further embodiment, provided herein arepharmaceutical compositions comprising 50-200 mg of a IL-7 bindingprotein which is an antibody comprising a light chain amino acidsequence as set out in SEQ ID NO:18 and a heavy chain amino acidsequence as set out in SEQ ID NO:19. In a further embodiment, providedherein are pharmaceutical compositions comprising 50-200 mg of a IL-7binding protein which is an antibody comprising a light chain amino acidsequence as set out in SEQ ID NO:20 and a heavy chain amino acidsequence as set out in SEQ ID NO:21. In a further embodiment, providedherein are pharmaceutical compositions comprising 50-200 mg of a IL-7binding protein which is an antibody comprising a light chain amino acidsequence as set out in SEQ ID NO:22 and a heavy chain amino acidsequence as set out in SEQ ID NO:23. In some embodiments, apharmaceutical composition described herein contains 30 mg, 31 mg, 32mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg 69 mg, 70 mg, 71 mg, 72 mg,73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, 100 mg, 101 mg, 102 mg,103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, or 110 mg of anIL-7 binding protein disclosed herein.

In some embodiments, the therapeutic agent of the disclosure (IL-7binding protein), when in a pharmaceutical preparation, is present inunit dose forms. In some embodiments, the dosage regimen will bedetermined by a medical profession and/or clinical factors. As is wellknown in the medical arts, dosages for any one patient depend upon manyfactors, including the patient's size, body surface area, age, thecompound to be administered, sex, time and route of administration,general health, and other drugs being administered concurrently.Exemplary doses can vary according to the size and health of theindividual being treated, as well as the condition being treated. Forexample, in some embodiments, the disclosed antibodies or functionalfragments may be administered in a dose of 1-100 mg/kg. In someembodiments, the disclosed IL-7 binding proteins may be administered ina dose of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or100 mg/kg. In some embodiments, a pharmaceutical composition disclosedherein is administered in a volume of at greater than at most about 5mL, 10 mL, 15 mL, 20 mL, 25 mL, 30 mL, 35 mL, 40 mL, 45 mL, 50 mL, 60mL, 70 mL, 80 mL, 90 mL, 100 mL, 110 mL, 120 mL, 130 mL, 140 mL, 150 mL,200 mL, 300 mL, 400 mL, or 500 mL. In some embodiments, pharmaceuticalcompositions disclosed herein are administered multiple times at thesedosages. In some embodiments, the dosage is administered a single timeor multiple times, for example daily, weekly, biweekly, or monthly,hourly, or is administered upon recurrence, relapse or progression of adisease or condition being treated. In some embodiments, administrationof a dose may be by slow continuous infusion over a period of from about2 to about 24 hours, such as from about 2 to about 12 hours, or fromabout 2 to about 6 hours.

In some embodiments, the pharmaceutical composition comprises acomposition for parenteral, transdermal, intraluminal, intraarterial,intrathecal and/or intranasal administration or by direct injection intotissue. In some embodiments, the pharmaceutical composition isadministered to a patient via infusion or injection. In one embodiment,provided are pharmaceutical compositions comprising an IL-7 bindingprotein for intravenous administration. In some embodiments, providedare pharmaceutical compositions comprising an IL-7 binding protein forsubcutaneous administration. In some embodiment, a pharmaceuticalcomposition described herein is administered to a subjecttransarterially, subcutaneously, intradermally, intratumorally,intranodally, intramedullary, intramuscularly, by intravenous (i.v.)injection, by intravenous (i.v.) infusion, or intraperitoneally. In someembodiments, the IL-7 binding protein or pharmaceutical compositionsthereof are administered to a subject by intradermal or subcutaneousinjection.

In some embodiments, a pharmaceutical composition is prepared by per seknown methods for the preparation of pharmaceutically acceptablecompositions that are administered to subjects, such that an effectivequantity of an IL-7 binding protein is combined in a mixture with apharmaceutically acceptable carrier. Suitable carriers are described,for example, in Remington's Pharmaceutical Sciences (Remington'sPharmaceutical Sciences, 20^(th) ed., Mack Publishing Company, Easton,Pa., USA, 2000). On this basis, the compositions may include, albeit notexclusively, solutions of the substances in association with one or morepharmaceutically acceptable carriers or diluents, and contained inbuffered solutions with a suitable pH and iso-osmotic with thephysiological fluids. In some embodiments, a pharmaceutical compositiondisclosed herein is acidic. In some embodiments, a pharmaceuticalcomposition disclosed herein is basic. In some embodiments, apharmaceutical composition can have a pH of about 1, 1.5, 2, 2.5, 3,3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5,12, 12.5, 13, 13.5, or about 14.

In some embodiments, suitable pharmaceutically acceptable carriersinclude essentially chemically inert and nontoxic compositions that donot interfere with the effectiveness of the biological activity of thepharmaceutical composition. Examples of suitable pharmaceutical carriersinclude, but are not limited to, water, saline solutions, glycerolsolutions, N-(1(2,3-dioleyloxy)propyl)N,N,N-trimethylammonium chloride(DOTMA), diolesylphosphotidyl-ethanolamine (DOPE), and liposomes. Insome embodiments, such compositions contain a therapeutically effectiveamount of an IL-7 binding protein disclosed herein, together with asuitable amount of carrier so as to provide the form for directadministration to a subject.

Pharmaceutical compositions may include, without limitation, lyophilizedpowders or aqueous or non-aqueous sterile injectable solutions orsuspensions, which may further contain antioxidants, buffers,bacteriostats and solutes that render the compositions substantiallycompatible with the tissues or the blood of an intended recipient. Othercomponents that may be present in such compositions include water,surfactants (such as Tween), alcohols, preservatives, polyols, glycerinand vegetable oils, for example. Extemporaneous injection solutions andsuspensions may be prepared from sterile powders, granules, tablets, orconcentrated solutions or suspensions.

A pharmaceutical composition disclosed herein may be formulated into avariety of forms and administered by a number of different means. Apharmaceutical formulation can be administered orally, rectally, orparenterally, in formulations containing conventionally acceptablecarriers, adjuvants, and vehicles as desired. The term “parenteral” asused herein includes subcutaneous, intravenous, intramuscular, orintrasternal injection and infusion techniques. Administration includesinjection or infusion, including intra-arterial, intracardiac,intracerebroventricular, intradermal, intraduodenal, intramedullary,intramuscular, intraosseous, intraperitoneal, intrathecal,intravascular, intravenous, intravitreal, epidural and subcutaneous),inhalational, transdermal, transmucosal, sublingual, buccal and topical(including epicutaneous, dermal, enema, eye drops, ear drops,intranasal, vaginal) administration. In some exemplary embodiments, aroute of administration is via an injection such as an intramuscular,intravenous, subcutaneous, or intraperitoneal injection.

Liquid formulations may include an oral formulation, an intravenousformulation, an intranasal formulation, an ocular formulation, an opticformulation, an aerosol, and the like. In certain embodiments, acombination of various formulations is administered. In certainembodiments a composition is formulated for an extended release profile.

Pharmaceutical compositions of the disclosure can be administered incombination with other therapeutics or treatments. In some embodiments,a treatment for a subject can be a surgery, a nutrition regime, aphysical activity, an immunotherapy, a pharmaceutical composition, acell transplantation, a blood fusion, or any combination thereof.

In some embodiments, a compositions/formulation disclosed herein is astable. In some embodiments, a “stable” formulation is one in which theIL-7 binding protein therein essentially retains its physical and/orchemical stability and/or biological activity upon storage. Variousanalytical techniques for measuring protein stability are available inthe art and are reviewed in Peptide and Protein Drug Delivery, 247-301,Vincent Lee Ed., Marcel Dekker, Inc., New York, N.Y., Pubs (1991) andJones, A. Adv. Drug Delivery Rev. 10: 29-90 (1993), for example.Stability can be measured at a selected temperature for a selected timeperiod. In some embodiments, the formulation is stable at ambienttemperature or at 40° C. for at least 1 month and/or stable at 2-8° C.for at least 1 to 2 years. In some embodiments, the formulation isstable following freezing (e.g. to −70° C.) and thawing. In someembodiments, a protein “retains its physical stability” in a formulationif it shows little to no change in aggregation, precipitation and/ordenaturation as observed by visual examination of color and/or clarity,or as measured by UV light scattering (measures visible aggregates) orsize exclusion chromatography (SEC). SEC measures soluble aggregatesthat are not necessarily a precursor for visible aggregates. In someembodiments, a protein “retains its chemical stability” in a formulationif the chemical stability at a given time is such that the protein isconsidered to retain its biological activity. Chemically degradedspecies may be biologically active and chemically unstable. Chemicalstability can be assessed by detecting and quantifying chemicallyaltered forms of the protein. Chemical alteration may involve sizemodification (e.g. clipping) which can be evaluated using SEC, SDS-PAGEand/or matrix-assisted laser desorption ionization/time-of-flight massspectrometry (MALDI/TOF MS), for example. Other types of chemicalalteration include charge alteration (e.g. occurring as a result ofdeamidation) which can be evaluated by ion-exchange chromatography, forexample.

In some embodiments, an IL-7 binding protein described herein is a dimerat a concentration of least about 2, 3, 4, 5 mg/ml in an acetate bufferand is monomeric at a concentration of less than about 5, 4, 3, 2, 1mg/ml. In some embodiments, an IL-7 binding protein described herein isa dimer at a concentration of about 5 mg/ml in an acetate buffer and ismonomeric at a concentration of about 1 mg/ml. In some embodiments, theIL-7 binding protein is a reversible dimer. In some embodiments, theIL-7 binding protein is a dimer. In some embodiments, the IL-7 bindingprotein is a monomer.

In an some embodiments, an IL-7 binding protein “retains its biologicalactivity” in a pharmaceutical formulation, if the biological activity ofthe IL-7 binding protein at a given time is within about 10% (within theerrors of the assay) of the biological activity exhibited at the timethe pharmaceutical formulation was prepared as determined in an antigenbinding assay, for example.

In some embodiments, a buffer disclosed herein refers to a bufferedsolution that resists changes in pH by the action of its acid-baseconjugate components. In some embodiments, a buffer can be phosphate,citrate and other organic acids. In some embodiments, a buffer isselected from the group consisting of sodium acetate, sodium carbonate,citrate, glycylglycine, histidine, glycine, lysine, arginine, sodiumdihydrogen phosphate, disodium hydrogen phosphate, sodium phosphate,sodium citrate, sodium borate, tris(hydroxymethyl)-aminomethan, bicine,tricine, malic acid, succinate, maleic acid, fumaric acid, tartaricacid, aspartic acid or mixtures thereof. A composition disclosed hereincan comprise antioxidants including ascorbic acid and/or methionine. Insome embodiments, a composition disclosed herein comprises apreservative. In some embodiments, a preservative is a compound whichcan be included in a formulation to essentially reduce microbialincluding bacterial action therein, thus facilitating the production ofa multi-use formulation, for example. Examples of potentialpreservatives include octadecyldimethylbenzyl ammonium chloride;hexamethonium chloride; benzalkonium chloride, benzethonium chloride;phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol);low molecular weight (less than about 10 residues) polypeptides;proteins, such as serum albumin, gelatin, or immunoglobulins;hydrophilic polymers such as polyvinylpyrrolidone; amino acids such asglycine, glutamine, asparagine, histidine, arginine, or lysine;monosaccharides, disaccharides, and other carbohydrates includingglucose, mannose, or dextrins; chelating agents such as EDTA; sugarssuch as sucrose, mannitol, trehalose or sorbitol; salt-formingcounter-ions such as sodium; metal complexes (e.g. Zn-proteincomplexes); and/or non-ionic surfactants such as TWEEN™, PLURONICS™ orpolyethylene glycol (PEG).

In some embodiments, a composition disclosed herein may further comprisea chemotherapeutic agent, cytotoxic agent, cytokine, growth inhibitoryagent, anti-hormonal agent, and/or cardioprotectant. Such molecules aresuitably present in combination in amounts that are effective for thepurpose intended.

In some embodiments, an IL-7 binding protein disclosed herein may beentrapped in microcapsules prepared, for example, by coacervationtechniques or by interfacial polymerization, for example,hydroxymethylcellulose or gelatin-microcapsules andpoly-(methylmethacylate) microcapsules, respectively, in colloidal drugdelivery systems (for example, liposomes, albumin microspheres,microemulsions, nano-particles and nanocapsules) or in macroemulsions.Such techniques are disclosed in Remington's Pharmaceutical Sciences16th edition, Osol, A. Ed. (1980).

In some embodiments, an IL-7 binding protein disclosed herein isprepared in a sustained-release preparation. Suitable examples ofsustained-release preparations include semipermeable matrices of solidhydrophobic polymers containing the antibody, which matrices are in theform of shaped articles, e.g. films, or microcapsules. Examples ofsustained-release matrices include polyesters, hydrogels (for example,poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides,copolymers of L-glutamic acid and γ ethyl-L-glutamate, non-degradableethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymerssuch as the LUPRON DEPOT™ (injectable microspheres composed of lacticacid-glycolic acid copolymer and leuprolide acetate), andpoly-D-(−)-3-hydroxybutyric acid.

In some embodiments, disclosed herein are pharmaceutical compositionscomprising the IL-7 binding protein which is present in a concentrationfrom 1 mg/ml to 500 mg/ml, and wherein the composition has a pH from 2.0to 10.0. The composition may further comprise a buffer system,preservative(s), tonicity agent(s), chelating agent(s), stabilizers andsurfactants. In some embodiments, the pharmaceutical composition is anaqueous formulation, for example, formulation comprising water. Suchformulation is typically a solution or a suspension. In a furtherembodiment, the pharmaceutical formulation is an aqueous solution. Insome embodiments, an aqueous formulation is a formulation comprising atleast 50% w/w water. In some embodiments an aqueous solution is definedas a solution comprising at least 50% w/w water. In some embodiments,the pharmaceutical composition is a stable liquid aqueous pharmaceuticalformulation comprising an anti-human IL-7 binding protein describedherein at a concentration of 20 to 150 mg/ml, a tonicity agent, asurfactant, and a buffer system having a pH of 4.0 to 8.0.

The pharmaceutical compositions may also comprise additional stabilizingagents, that may further enhance stability of a therapeutically activeIL-7 binding protein. Stabilizing agents of can include, but are notlimited to, methionine and EDTA, which protect the polypeptide againstmethionine oxidation, and a nonionic surfactant, which protects thepolypeptide against aggregation associated with freeze-thawing ormechanical shearing. In some embodiments, the composition may furthercomprise a surfactant. The surfactant may be selected from a detergent,ethoxylated castor oil, polyglycolyzed glycerides, acetylatedmonoglycerides, sorbitan fatty acid esters,polyoxypropylene-polyoxyethylene block polymers (e.g. poloxamers such asPLURONIC F68, poloxamer 188 and 407, Triton X-100), polyoxyethylenesorbitan fatty acid esters, polyoxyethylene and polyethylene derivativessuch as alkylated and alkoxylated derivatives (tweens, e.g. Tween-20,Tween-40, Tween-80 and Brij-35), monoglycerides or ethoxylatedderivatives thereof, diglycerides or polyoxyethylene derivativesthereof, alcohols, glycerol, lectins and phospholipids (eg. phosphatidylserine, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidylinositol, diphosphatidyl glycerol and sphingomyelin), derivates ofphospholipids (eg. dipalmitoyl phosphatidic acid) and lysophospholipids(eg. palmitoyl lysophosphatidyl-L-serine andI-acyl-sn-glycero-3-phosphate esters of ethanolamine, choline, serine orthreonine) and alkyl, alkoxyl (alkyl ester), alkoxy (alkylether)-derivatives of lysophosphatidyl and phosphatidylcholines, e.g.lauroyl and myristoyl derivatives of lysophosphatidylcholine,dipalmitoylphosphatidylcholine, and modifications of the polar headgroup, that is cholines, ethanolamines, phosphatidic acid, serines,threonines, glycerol, inositol, and the positively charged DODAC, DOTMA,DCP, BISHOP, lysophosphatidylserine and lysophosphatidylthreonine, andglycerophospholipids (eg. cephalins), glyceroglycolipids (eg.galactopyransoide), sphingoglycolipids (eg. ceramides, gangliosides),dodecylphosphocholine, hen egg lysolecithin, fusidic acidderivatives—(e.g. sodium tauro-dihydrofusidate etc.), long-chain fattyacids and salts thereof C6-C12 (eg. oleic acid and caprylic acid),acylcarnitines and derivatives, N^(a)-acylated derivatives of lysine,arginine or histidine, or side-chain acylated derivatives of lysine orarginine, N^(a)-acylated derivatives of dipeptides comprising anycombination of lysine, arginine or histidine and a neutral or acidicamino acid, N^(a)-acylated derivative of a tripeptide comprising anycombination of a neutral amino acid and two charged amino acids, DSS(docusate sodium, CAS registry no [577-11-7]), docusate calcium, CASregistry no [128-49-4]), docusate potassium, CAS registry no[7491-09-0]), SDS (sodium dodecyl sulphate or sodium lauryl sulphate),sodium caprylate, cholic acid or derivatives thereof, bile acids andsalts thereof and glycine or taurine conjugates, ursodeoxycholic acid,sodium cholate, sodium deoxycholate, sodium taurocholate, sodiumglycocholate, N-Hexadecyl-N,N-dimethyl-3-ammonio-I-propanesulfonate,anionic (alkyl-aryl-sulphonates) monovalent surfactants, zwitterionicsurfactants (e.g. N-alkyl-N,N-dimethylammonio-I-propanesulfonates,3-cholamido-I-propyldimethylammonio-I-propanesulfonate, cationicsurfactants (quaternary ammonium bases) (e.g. cetyl-trimethylammoniumbromide, cetylpyridinium chloride), non-ionic surfactants (eg. Dodecylβ-D-glucopyranoside), poloxamines (eg. Tetronic's), which aretetrafunctional block copolymers derived from sequential addition ofpropylene oxide and ethylene oxide to ethylenediamine, or the surfactantmay be selected from the group of imidazoline derivatives, or mixturesthereof.

Diagnostic/Monitoring

In some embodiments, an IL-7 binding protein disclosed herein is used ina method of diagnosis or prognosis. In some embodiments, diagnosisincludes determining whether a subject has a disease or condition and/ordetermining the severity of the disease or condition. In someembodiments, prognosis includes predicting whether or not a subject willdevelop a disease or condition, whether or not they will need treatment,the type of treatment the individual will need, whether or not they willrespond to a treatment, whether or not and/or when they will suffer adisease episode, recurrence or relapse, and the severity or duration ofa symptom or a disease episode, recurrence or relapse. In someembodiments, a method of diagnosis or prognosis may include selecting orrecommending a suitable treatment for the individual, for example, basedon the diagnosis or prognosis. In some embodiments, a selected orrecommended treatment or combination of treatments may then beadministered to the subject.

In some embodiments, an IL-7 binding protein disclose herein is used todiagnose or use in prognosis of an autoimmune or inflammatory disease,particularly those in which pathogenic T_(H)17 cells are implicated.Such diseases are associated with high levels of IL-17 expression.Elevated levels of IL-17 have been reported in serum and CSF of MSpatients (Matusevicius, D. et al.; Mult. Scler. 5, 101-104; 1999) and inthe synovial fluid obtained from rheumatoid arthritis patients. IL-17has also been implicated in psoriasis (Homey et al.; J. Immunol.164(12):6621-32; 2000), while Hamzaoui et al reported high levels ofIL-17 in Behcet's disease (Scand. J. Rhuematol.; 31:4, 205-210; 2002).Elevated IL-17 levels have also been observed in systemic lupuserythematosus (SLE) (Wong et al.; Lupus 9(8):589-93; 2000).

In some embodiments, a method disclosed herein comprises measuring alevel of IL-7 in a subject or on/in a sample obtained from a subject. Insome embodiments, measuring a level of IL-7 can be performed by a methodand assay known in the art. In some embodiments, a level of IL-7 iscompared to a reference level of IL-7. In some embodiments, a referenceslevel is indicative of a normal, non-diseases, disease, or diseasestage. In some embodiments, a level of IL-7 is measured multiple timesin a subject or from multiple samples obtained from a subject. In someembodiment, diagnostic or prognostic methods can be carried out inconjunction with one or more other assays or tests to refine thediagnosis or prognosis. For example, other markers may be included inthe analysis.

Assaying a tissue sample of a subject may be performed at one or moretime points. A separate tissue sample may be obtained from the subjectfor assaying at each of the one or more time points. Assaying at one ormore time points may be performed on the same tissue sample. Assaying atone or more time points may provide an assessment of an effectiveness ofa drug, a longitudinal course of a disease treatment regime, or acombination thereof. At each of the one or more time points, a tissuesample may be compared to a same reference. A tissue sample may becompared to a different reference at each of the one or more timepoints. The one or more time points may be the same. The one or moretime points may be different. The one or more time points may compriseat least one time point prior to a therapeutic administration, at leastone time point after a therapeutic administration, at least one timepoint prior to a positive disease diagnosis, at least one time pointafter a disease remission diagnosis, at least one time point during adisease treatment regime, or a combination thereof.

The methods as described herein may be used to monitor a subject havingrisk of developing a disease or condition, as a preventive measure. Themethods as described herein may be used alone for diagnosis and/ormonitoring efficacy of a treatment. The methods as described herein maybe used in combination with other assays for diagnosis or monitoring(such as a cytological analysis or molecular profiling).

In some embodiments, an increased level of IL-7 or IL-7 receptor in asubject or sample, as compared with a reference sample or referencelevel, indicates a positive diagnosis relating to the presence ofdisease, for example that the individual has the relevant disease orcondition or has more severe disease. In some embodiments, a subjecthaving a disease or condition includes an individual suspected of havingthe disease or condition and/or an individual at risk of developing thedisease or condition. For example, the individual may not have beenformally diagnosed but may be suspected of having the disease orcondition because of the presence of one or more symptoms. For example,IL-7 expression is increased in the circulation of individuals withlymphopenia and there is a strong inverse correlation betweencirculating IL-7 levels and the number of CD4⁺ T cells (Mackall et al.,2011). In some embodiment, an IL-7 binding protein disclosed herein isused to detect lymphopenia.

In some embodiments, a method for predicting the responsiveness of asubject to a treatment may be carried out before administration atherapy. The prediction may then be taken into account when selecting orrecommending a suitable treatment for the individual. Alternatively, amethod of predicting responsiveness to a treatment may be carried outafter treatment with a therapy and used to monitor and predict thesubject's response to treatment. For example, systemic levels of IL-7may increase with the duration of disease (Khaibullin et al., 2017) andincreased circulating IL-7 has been identified as a potential predictivebiomarker for response to IFN-3 treatment in MS. In some embodiments,circulating IL-7 increases in Chron's and ulcerative colitis patients aswell as patients with SLE. In some embodiments, a disease or conditiondisclosed herein can be detected in a sample based on a level of IL-7 inthe sample.

A sample obtained from a subject can comprise tissue, cells, cellfragments, cell organelles, nucleic acids, genes, gene fragments,expression products, gene expression products, gene expression productfragments or any combination thereof. A sample can be heterogeneous orhomogenous. A sample can comprise blood, urine, cerebrospinal fluid,seminal fluid, saliva, sputum, stool, lymph fluid, tissue, mucus, or anycombination thereof. A sample can be a tissue-specific sample such as asample obtained from a reproductive tissue (such as a sperm or an egg),thyroid, skin, heart, lung, kidney, breast, pancreas, liver, muscle,smooth muscle, bladder, gall bladder, colon, intestine, brain,esophagus, prostate, or any combination thereof. In some embodiments, asample can be a cell-free sample.

As used herein, the term “cell-free” refers to the condition of thenucleic acid sequence as it appeared in the body before the sample isobtained from the body. For example, circulating cell-free nucleic acidsequences in a sample may have originated as cell-free nucleic acidsequences circulating in the bloodstream of the human body. In contrast,nucleic acid sequences that are extracted from a solid tissue, such as abiopsy, are generally not considered to be “cell-free.” In some cases,cell-free DNA may comprise fetal DNA, maternal DNA, or a combinationthereof. In some cases, cell-free DNA may comprise DNA fragmentsreleased into a blood plasma. In some cases, cell-free DNA may comprisecirculating DNA indicative of a tissue origin, a disease or a condition.A cell-free nucleic acid sequence may be isolated from a blood sample. Acell-free nucleic acid sequence may be isolated from a plasma sample.

A sample may be obtained from a subject by another individual or entity,such as a healthcare (or medical) professional or robot. A medicalprofessional can include a physician, nurse, medical technician orother. In some cases, a physician may be a specialist, such as anoncologist, surgeon, or endocrinologist. A medical technician may be aspecialist, such as a cytologist, phlebotomist, radiologist,pulmonologist or others. A medical professional may obtain a sample froma subject for testing or refer the subject to a testing center orlaboratory for the submission of the sample. The medical professionalmay indicate to the testing center or laboratory the appropriate test orassay to perform on the sample, such as methods of the presentdisclosure including determining gene sequence data, gene expressionlevels, sequence variant data, or any combination thereof. In somecases, a medical professional need not be involved in the initialdiagnosis of a condition or a disease or the initial sample acquisition.An individual, such as the subject, may alternatively obtain a samplethrough the use of an over the counter kit. The kit may containcollection unit or device for obtaining the sample as described herein,a storage unit for storing the sample ahead of sample analysis, andinstructions for use of the kit.

Kits

A kit-of-parts comprising a pharmaceutical composition together withinstructions for use is further provided. For convenience, thekit-of-parts may comprise reagents in predetermined amounts withinstructions for use.

In some embodiments, disclosed herein are kids comprising an IL-7binding protein disclosed herein. In some embodiments, a kit can be adiagnostic kit. In some embodiments, a kit comprises an IL-7 bindingprotein disclosed herein and instructions for use. In some embodiments,a kit comprises means for measuring IL-7 level in a sample andinstructions for use. A kit may provide a unit or device for obtaining asample from a subject (e.g., a device with a needle coupled to anaspirator). A kit may include a plurality of syringes, ampules, foilpackets, or blister packs, each containing a single unit dose of a kitcomponent described herein. Containers of a kit may be airtight,waterproof (e.g., impermeable to changes in moisture or evaporation),and/or light-tight. A kit may include a device suitable foradministration of the component, e.g., a syringe, inhalant, pipette,forceps, measured spoon, dropper (e.g., eye dropper), swab (e.g., acotton swab or wooden swab), or any such delivery device. In someembodiments, the device may be a medical implant device, e.g., packagedfor surgical insertion. A kit disclosed herein may comprise one or morereagents or instruments which enable the method to be carried out. Insome embodiments, reagents or instruments include one or more of thefollowing: suitable buffer(s) (aqueous solutions) a support comprisingwells on which quantitative reactions can be done. A kit may be aspecific kit for a specific tissue sample. Further, a kit disclosedherein may comprise a control.

In addition to the above components, instructions for use may beprovided in a kit. These instructions may be present in the kit in avariety of forms, such as printed information on a suitable medium orsubstrate (e.g., a piece or pieces of paper on which the information isprinted), in the packaging of the kit, in a package insert, etc. In someembodiments, instructions for use can be provided on a computer readablemedium (e.g., jump/thumb drive, CD, etc.), on which the information hasbeen recorded or at a website address which may be used via the internetto access the information at a website.

Devices

Another aspect of the disclosure provides a pre-filled syringe orautoinjector device, comprising an IL-7 binding protein or a compositiondescribed herein. In some embodiments, a composition stored in acontainer, pre-filled syringe, injector or autoinjector device containsan IL-7 binding protein disclosed herein.

Arrays

Disclosed herein are supports comprising an IL-7 binding proteindisclosed herein. A support can be a solid support. A support may take avariety of configurations ranging from simple to complex, depending onthe intended use of the support. A support can have an overall slide orplate configuration, such as a rectangular or disc configuration. Astandard microplate configuration can be used. In some embodiments, thesurface may be smooth or substantially planar, or have irregularities,such as depressions or elevations. In some embodiments, a support mayhave a rectangular cross-sectional shape, having a length of from about10-200 mm, 40-150 mm, or 75-125 mm; a width of from about 10-200 mm,20-120 mm, or 25-80 mm, and a thickness of from about 0.01-5.0 mm, 0.1-2mm, or 0.2 to 1 mm.

In some embodiments, a support can be organic or inorganic; may be metal(e.g., copper or silver) or non-metal; may be a polymer or nonpolymer;may be conducting, semiconducting or nonconducting (insulating); may bereflecting or nonreflecting; may be porous or nonporous; etc. A supportas described herein can be formed of any suitable material, includingmetals, metal oxides, semiconductors, polymers (particularly organicpolymers in any suitable form including woven, nonwoven, molded,extruded, cast, etc.), silicon, silicon oxide, and composites thereof. Asupport can be an array. In some embodiments, a support comprises anarray. An array can comprise an ordered spatial arrangement of two ormore discrete regions. An array can comprise IL-7 binding proteinslocated at known or unknown discrete regions. Row and columnarrangements of arrays can be selected due to the relative simplicity inmaking such arrangements. The spatial arrangement can, however, beessentially any form selected by the user, and optionally, in a pattern.Areas of an array may be any convenient shape, including circular,ellipsoid, oval, annular, or some other analogously curved shape, wherethe shape may, in certain embodiments, be a result of the particularmethod employed to produce the array.

In some embodiments, a support can be planar. In some instances, asupport can be spherical. In some instances, a support can be a bead. Insome instances, a support can be magnetic. In some embodiments, amagnetic support can comprises magnetite, maghemitite, FePt, SrFe, iron,cobalt, nickel, chromium dioxide, ferrites, or mixtures thereof. In someembodiments, a support can be nonmagnetic. In some embodiments, thenonmagnetic support can comprise a polymer, metal, glass, alloy,mineral, or mixture thereof. In some instances, a nonmagnetic materialcan be a coating around a magnetic support. In some instances, amagnetic material may be distributed in the continuous phase of amagnetic material. In some embodiments, the support comprises magneticand nonmagnetic materials. In some instances, a support can comprise acombination of a magnetic material and a nonmagnetic material. In someembodiments, an IL-7 binding protein disclosed herein is directly orindirectly associated with a support disclosed herein.

Computer Control Systems

The present disclosure provides computer control systems that areprogrammed to implement methods of the disclosure. In some embodiments,a computer system is programmed or otherwise configured to interfacewith an apparatus that is configured to detect IL-7 and/or binding of anIL-7 binding protein disclosed herein to moiety. The computer system canbe an electronic device of a user or a computer system that is remotelylocated with respect to the electronic device. The electronic device canbe a mobile electronic device.

In some embodiments, a computer system includes a central processingunit (CPU, also “processor” and “computer processor” herein, which canbe a single core or multi core processor, or a plurality of processorsfor parallel processing. In some embodiments, a computer system alsoincludes memory or memory location (e.g., random-access memory,read-only memory, flash memory), electronic storage unit (e.g., harddisk), communication interface (e.g., network adapter) for communicatingwith one or more other systems, and peripheral devices, such as cache,other memory, data storage and/or electronic display adapters. In someembodiments, the memory, storage unit, interface and peripheral devicesare in communication with the CPU through a communication bus, such as amotherboard. In some embodiments, the storage unit can be a data storageunit (or data repository) for storing data. In some embodiments, thecomputer system is operatively coupled to a computer network (“network”)with the aid of the communication interface. In some embodiments, thenetwork can be the Internet, an internet and/or extranet, or an intranetand/or extranet that is in communication with the Internet. In someembodiments, the network is a telecommunication and/or data network. Thenetwork can include one or more computer servers, which can enabledistributed computing, such as cloud computing. In some embodiments, thenetwork, in some cases with the aid of the computer system can implementa peer-to-peer network, which may enable devices coupled to the computersystem to behave as a client or a server. In some embodiments, the CPUexecutes a sequence of machine-readable instructions, which can beembodied in a program or software. The instructions may be stored in amemory location, such as the memory. The instructions can be directed tothe CPU, which can subsequently program or otherwise configure the CPUto implement methods of the present disclosure. Examples of operationsperformed by the CPU can include fetch, decode, execute, and writeback.In some embodiments, the CPU can be part of a circuit, such as anintegrated circuit. One or more other components of the system can beincluded in the circuit. In some cases, the circuit is an applicationspecific integrated circuit (ASIC). In some embodiments, the storageunit can store files, such as drivers, libraries and saved programs. Thestorage unit can store user data, e.g., user preferences and userprograms. The computer system in some cases can include one or moreadditional data storage units that are external to the computer system,such as located on a remote server that is in communication with thecomputer system through an intranet or the Internet. In someembodiments, the computer system communicates with one or more remotecomputer systems through the network. For instance, the computer systemcan communicate with a remote computer system of a user. Examples ofremote computer systems include personal computers (e.g., portable PC),slate or tablet PC's telephones, Smart phones, or personal digitalassistants. The user can access the computer system via the network.

Methods as described herein can be implemented by way of machine (e.g.,computer processor) executable code stored on an electronic storagelocation of the computer system, such as, for example, on the memory orelectronic storage unit. The machine executable or machine-readable codecan be provided in the form of software. During use, the code can beexecuted by the processor. In some cases, the code can be retrieved fromthe storage unit and stored on the memory for ready access by theprocessor. In some situations, the electronic storage unit can beprecluded, and machine-executable instructions are stored on memory. Insome embodiments, the code can be pre-compiled and configured for usewith a machine having a processer adapted to execute the code, or can becompiled during runtime. The code can be supplied in a programminglanguage that can be selected to enable the code to execute in apre-compiled or as-compiled fashion.

Aspects of the systems and methods provided herein, such as the computersystem, can be embodied in programming. Various aspects of thetechnology may be thought of as “products” or “articles of manufacture”typically in the form of machine (or processor) executable code and/orassociated data that is carried on or embodied in a type of machinereadable medium. Machine-executable code can be stored on an electronicstorage unit, such as memory (e.g., read-only memory, random-accessmemory, flash memory) or a hard disk. “Storage” type media can includeany or all of the tangible memory of the computers, processors or thelike, or associated modules thereof, such as various semiconductormemories, tape drives, disk drives and the like, which may providenon-transitory storage at any time for the software programming. All orportions of the software may at times be communicated through theInternet or various other telecommunication networks. Suchcommunications, for example, may enable loading of the software from onecomputer or processor into another, for example, from a managementserver or host computer into the computer platform of an applicationserver. Thus, another type of media that may bear the software elementsincludes optical, electrical and electromagnetic waves, such as usedacross physical interfaces between local devices, through wired andoptical landline networks and over various air-links. The physicalelements that carry such waves, such as wired or wireless links, opticallinks or the like, also may be considered as media bearing the software.As used herein, unless restricted to non-transitory, tangible “storage”media, terms such as computer or machine “readable medium” refer to anymedium that participates in providing instructions to a processor forexecution.

A machine readable medium, such as computer-executable code, may takemany forms, including but not limited to, a tangible storage medium, acarrier wave medium or physical transmission medium. Non-volatilestorage media include, for example, optical or magnetic disks, such asany of the storage devices in any computer(s) or the like, such as maybe used to implement the databases, etc. shown in the drawings. Volatilestorage media include dynamic memory, such as main memory of such acomputer platform. Tangible transmission media include coaxial cables;copper wire and fiber optics, including the wires that comprise a buswithin a computer system. Carrier-wave transmission media may take theform of electric or electromagnetic signals, or acoustic or light wavessuch as those generated during radio frequency (RF) and infrared (IR)data communications. Common forms of computer-readable media thereforeinclude for example: hard disk, magnetic tape, any other magneticmedium, a CD-ROM, DVD or DVD-ROM, any other optical medium, punch cardspaper tape, any other physical storage medium with patterns of holes, aRAM, a ROM, a PROM and EPROM, a FLASH-EPROM, any other memory chip orcartridge, a carrier wave transporting data or instructions, cables orlinks transporting such a carrier wave, or any other medium from which acomputer may read programming code and/or data. Many of these forms ofcomputer readable media may be involved in carrying one or moresequences of one or more instructions to a processor for execution.

In some embodiments, a computer system disclosed herein can include orbe in communication with an electronic display that comprises a userinterface (UI) for providing, for example, one or more results(immediate results or archived results from a previous experiment), oneor more user inputs, reference values from a library or database, or acombination thereof. Examples of UIs include, without limitation, agraphical user interface (GUI) and web-based user interface.

Further, methods and systems of the present disclosure can beimplemented by way of one or more algorithms. An algorithm can beimplemented by way of software upon execution by the central processingunit. The algorithm can, for example, determine optimized conditions viasupervised learning to optimize conditions such as a buffer type, abuffer concentration, a temperature, an incubation period, thresholds,diagnostic/prognostic indications for methods disclosed herein.

EXAMPLES Example 1: DRSPAI-L7B In Cynomolgus Monkey

The study was set forth to evaluate DRSPAI-L7B in cynomolgus monkey.Part 1, three single dose IV injections at 0.1, 1 and 10 mg/kg wereevaluated. In part 2, repeat dosing (four SC injections) of 30 mg/kg inthe vehicle control animals from part 1. Blood samples were collectedfor assessment of drug PK as well as free and total IL-7 levels andpharmacodynamic activity of DRSPAI-L7B through the measurement of STAT5phosphorylation.

Total antibody levels Total antibody levels were determined using ageneric antigen capture and detection method on the GYROLAB. The maximumdetected peak serum concentration (C_(max)) and the time at which it wasobserved (T_(m)ax) were determined by inspection of the obtained data.In addition, the AUC, total serum clearance (CL); volume of distributionat steady-state (Vss) and terminal half-life (t½) were calculated usinga non-compartmental PK model. (Table 3)

TABLE 3 Mean PK parameters for DRSPAI-L7B (IV administration) for singleascending dose part 1 and repeat dose part 2 Part 1 Dose C_(max)AUC_(inf) Half-Life CL V_(ss) (mg/kg) (ug/mL) (hr*ug/mL) (hr) (mL/hr/Kg)(mL/Kg) 0.1 3.12 721 350 0.14 82 (2.80- (710- (270- (0.14- (71- 3.55)733) 410) 0.14) 90) 1.0 37.2 8250 470 0.12 71 (27.8- (7520- (350- (0.11-(58- 48.4) 9060) 560) 0.13) 80) 10.0 307 93100 420 0.11 66 (262- (76800-(340- (0.09- (58- 337) 112000) 490) 0.13) 73) Part 2 (30 mg/kg) C_(max)AUC_(i0-168) Period (ug/mL) (hr*ug/mL) First dose 320 41600 (258-398)(32700-47300) Second dose 463 N/A (168 h post dose) (407-516) Third dose556 N/A (168 h post dose) (490-602) Fourth dose 911 119000 (688-1270)(94100-169000)

No target mediated drug disposition (TMDD) was observed and as the doseincreases from 0.1 to 10 mg/kg the increase in serum Cmax and AUC aredose proportional. DRSPAI-L7B is cleared slowly with a mean half-life ofapproximately 17±3.9 days and on repeat dosing, a 2.8-fold accumulationof drug. The data is presented in FIG. 1A, FIG. 1B, and FIG. 1C.

An acid dissociation bridging assay was used to assess the presence ofanti-drug antibodies (ADA) using DRSPAI-L7B to capture. No ADA weredetected during either part 1 or part 2 of the study.

A further study was carried out to determine the PK and PD of DRSPAI-L7Bin cynomolgus monkeys following subcutaneous (SC) dosing. Four singledose IV injections at 0.1, 1, 3 and 10 mg/kg were evaluated. The doseswere administered by SC injection once on Day 1.

The systemic exposure to DRSPAI-L7B was determined by calculating thearea under the serum concentration time curve (AUC) from the start ofdosing to the last quantifiable time point (AUC_(0-t)) using the linearup/log down trapezoidal method. The maximum observed peak serumconcentration (Cmax) and the time at which it was observed (Tmax) weredetermined by PK SUBMIT.

TABLE 4 Serum PK parameters for DRSPAI-L7B (SC administration) Dose ofDRSPAI-L7B (mg/kg) Parameter 0.1 1 3 10 AUC_(0-t) Mean 943 6960 1890060200 (μg · h/ml) Min 687 3870 17100 52700 Max 1120 9130 21300 69700C_(max) Mean 2.01 18.1 34.2 119 (μg/ml) Min 1.76 16.9 29.5 108 Max 2.2119.0 39.1 128 T_(max) Mean 168 96 96 96 (h) Min 96 48 96 96 Max 168 9696 168

At the highest dose, 10 mg/kg, the gender averaged mean Cmax was 119Dg/mL (range from 108 to 128 μg/mL), and mean AUC_(0-t) 60200 μg·h/mL(range from 52700 to 69700 μg·h/mL).

There were dose-dependent increases in total IL-7 levels at all doselevels, demonstrating target engagement of DRSPAI-171B. There was alsodose-dependent inhibition of IL-7 induced STAT5 phosphorylation in totalTh and Tc lymphocytes at 1 mg/kg and a dose dependent decrease in Bcl-2expression in Th lymphocytes at ≥3 mg/kg.

Low levels of anti-DRSPAI-171B antibodies were detected in two of threemonkeys given 1.0 mg/kg. In one male this resulted in reduced targetengagement and a lower AUC.

Example 2: DRSPAI-L7B Binding Affinity to IL-7

The kinetics and affinities for binding of DRSPAI-171B to human andcynomolgus monkey IL-7 were assessed at 25° C. and 37° C. by surfaceplasmon resonance (SPR) using a Biacore 8K instrument. The affinity ofDRSPAI-171B for human IL-7 was approximately 34 pM at 25° C. and 67 pMat 37° C. (Table 5). The affinity of DRSPAI-17B for cynomolgus IL-7 wasapproximately 53 pM at 25° C. and 75 pM at 37° C. (Table 5).

TABLE 5 Binding kinetics and affinities of DRSPAI- L7B to human andcynomolgus IL-7 average ka average kd Average KD SD Analyte (1/Ms) (1/s)(pM) (KD, pM) DRSPAI-L7B at 25° C. hIL-7 6.49E+06 2.06E−04 31 5 hIL-74.65E+06 1.58E−04 34 7 cynoIL-7 3.17E+06 1.70E−04 53 6 DRSPAI-L7B at 37°C. hIL-7 8.28E+06 5.68E−04 69 5 hIL-7 1.47E+07 6.78E−04 46 8 hIL-71.10E+07 8.26E−04 75 12 hIL-7 5.90E+06 5.15E−04 87 3 hIL-7: geometricmean KD (pM)* 67.4 (58.4, 76.3) cynoIL-7 6.17E+06 4.62E−04 73 5*geometric mean KD derived from all experiments (pM, with 85% confidenceinterval)

Example 3: Inhibition of IL-7 Signaling—Functional Assay

All human samples were obtained with patient informed consent inaccordance with ICH GCP under a protocol approved by a national,regional or investigational center ethics committee or an InstitutionalReview Board (IRB) approved protocol. Disease PBMCs were supplied by anapproved external human tissue supplier. PBMCs were stored frozen at−80° C. until use.

Healthy volunteer blood was provided by a Blood Donation Unit: Blood waswithdrawn by venepuncture and transferred into a pot or blood bagcontaining sodium heparin (1 U/mL). The blood was collected and usedwithin 1 hour, either for whole blood assays or for PBMC isolation.Different donors were used for each experiment. Cells were thawed byremoving from −80° C. storage and immediately placed into a water bathat 37° C. After transferring cell suspension to a 15 mL centrifuge tubewarm medium (RPMI+10% heat-inactivated FCS, 1% penicillin/streptomycin,and 1% GlutaMax) was added very slowly to decrease DMSO concentrationgradually. Once volume was increased to 10 mL the cells were centrifugedand washed once more before counting and resuspending in an appropriatevolume of assay medium to yield 5×10⁶ cells per 1 mL.

All antibodies were aliquoted on arrival for long term storage at −80°C. For experimentation, antibody aliquots were thawed and stored at 4°C. for no longer than 8 weeks.

Reagent Supplier AIM V Medium Gibco Recombinant human IL-7 R&D SystemsBD Phosflow ™ Lyse/Fix Buffer 5x BD Bioscience BD Phosflow ™ Perm BufferIII BD Biosciences Flow cytometry staining buffer eBioscience FcRBlocking reagent Miltenyi Biotec BD Phosflow ™ PE Mouse anti-STAT5(pY694) BD Bioscience Mouse anti-human CD8 FITC (clone: SKI) BiolegendMouse anti-human CD4 PerCP/Cy5.5 (clone: RPA-T4) Biolegend Mouseanti-human CD3 BV510 (clone: SK7) Biolegend Anti-mouse Ig, κ/NegativeControl Compensation BD Bioscience Particles Set BD FACSDiva CS&TResearch Beads BD Bioscience

All antibody treatments and rhIL-7 stimulations were made up at 4× finalassay concentration (F.A.C.) in culture medium before mixing 1:1 andincubating at room temperature for 5 minutes. In the absence of eitherantibody or IL-7 stimulus, culture medium was added.

For whole blood: 100 μL of the antibody:IL-7 mixture was aliquoted intoFACS tubes before adding 100 μL of whole blood. Tubes were mixed gentlyby vortexing and incubated at 37° C. in a humidified incubator for 20minutes. At the end of the stimulation period 2.5 mL pre-warmed PHOSFLOWlysis buffer (1×) was added and the samples were incubated for a further10 minutes at 37° C. 2 mL PBS was added to the suspension and tubes werecentrifuged to pellet the cells (300×g, 5 minutes at room temperature).Supernatant was discarded and cells were washed twice more in PBS.Following the final wash, cells were resuspended in 500 μL Perm BufferIII (pre-cooled to −20° C.) and vortexed to mix. Tubes were incubated onice for 30 minutes before washing once in 3 mL PBS. Cells wereresuspended in 100 μL PBS and transferred to 96-well, round bottomculture plate for staining.

For PBMCs treatments: 50 μL of the antibody:IL-7 mixture was added toeach well of a 96-well, round bottom tissue culture plate. 50 μL PBMCsuspension was added to each treatment well (2.5×10⁵ cells/well). Plateswere mixed gently on a rotary plate shaker before incubating at 37° C.in a humidified incubator for 20 minutes. At the end of the stimulationperiod 250 μL pre-warmed PHOSFLOW lysis buffer (1×) was added and thesamples were incubated for a further 10 minutes at 37° C. Followingfixation, cells were pelleted by centrifugation (300×g, 5 minutes atroom temperature) and washed twice in 200 μL PBS. Cell pellets wereresuspended in 100 μL Perm Buffer III (pre-cooled to −20° C.) andpipetted up and down to mix. Cells were incubated on ice for 30 minutesbefore washing once in 200 μL PBS and resuspending in PBS.

For all samples: Following permeabilization, plates were centrifuged(300×g, 5 minutes at room temperature) and cell pellets were resuspendedin 25 μL FcR blocking reagent diluted 1:5 in 3% BSA in PBS. Cells wereincubated for 10 minutes at room temperature before adding 2.5 μLanti-CD3 (BV510), 2.5 μL anti-CD4 (PerCP/Cy5.5), 2.5 anti-CD8 (FITC),7.5 μL anti-pSTAT5 (PE) and 35 μL flow cytometry staining buffer (50 μLtotal per sample). Where a stain was excluded for controls, theequivalent volume of staining buffer was added instead. Plates weremixed briefly on a rotary plate shaker and incubated on ice for 30minutes protected from light. Cells were washed in 200 μL stainingbuffer and resuspended in 200 μL staining buffer for analysis on thesame day on the FACS Canto II.

The performance of the instrument was checked using the Cytometer Set-upand Tracking (CST) Beads. This is a QC check for the instrument, setsthe baselines and optimizes the voltages for each laser prior to use.The results of the calibration are stored within the CST software on theinstrument.

Compensation for the instrument was performed using anti-mouseIgG,_(κ)/negative control compensation beads in accordance with themanufacturer's instructions. The relevant antibodies used to stain thecells during the experiment were used to label the appropriatecompensation bead type. Compensation for the experiments were performedusing the appropriately labelled beads with the automatic compensationfacility available within the FACS Diva software. After analysis of thecompensation samples the appropriate compensation settings werecalculated and applied to each experimental staining panel. pSTAT5 FACSanalysis: Acquired cells in a FSC-A vs FSC-H plot to exclude doublets.Single cells were acquired into a FSC-A vs SSC-A plot and gated aroundthe live lymphocytes. These were acquired into AmCyan vs FSC-A plot andgated around the CD3⁺ population. A PerCP-Cy5.5 vs FITC plot wasgenerated on CD3⁺ to identify CD4⁺ and CD8⁺ populations. For pSTAT5 ahistogram for PE fluorescence was created for each subset. Positivegates were set based on the unstimulated samples and the percent PEpositive statistic was used for data analysis.

Data were analyzed using FlowJo software (version 10) and results weregenerated in Microsoft Excel (2010) spreadsheet format using the BatchAnalysis facility within the FlowJo software. The cell populations weretabulated in an Excel spreadsheet as % of parent. This was convertedinto a percent response by normalizing to the unstimulated control or apercent inhibition by normalizing to the unstimulated control andsubtracting this value from 100 (theoretical maximal percent response).

Where concentration response graphs have been generated, individualdonor data were fitted with a non-linear logistic curve fit regressionand the average (mean and median), range, SD and SEM of the IC₅₀ fromall donors was calculated in GraphPad Prism (version 6). Wherestatistical testing was carried out, a 2-way ANOVA with Sidak'scorrection for multiple comparison was applied. A p value of <0.05 wasconsidered as statistically significant. * denotes p<0.05; **p≤0.01;***p≤0.001; and ****p<0.0001.

In a whole blood assay, STAT5 phosphorylation was assessed in CD4⁺ Tcells by flow cytometry after 20 minutes stimulation with 1 ng/mlrecombinant human IL-7 (rhIL-7; 58 pM). As shown in FIG. 2A and Table 6,DRSPAI-L7B prevented IL-7 from signaling through STAT5 in aconcentration-dependent manner with a median IC₅₀ of 34 pM (5.1 ng/mL).A1290 prevented IL-7 signalling through STAT5 in aconcentration-dependent manner with a median IC₅₀ of 18 pM (0.00275μg/ml). A1291 prevented IL-7 signalling through STAT5 in aconcentration-dependent manner with a median IC₅₀ of 16 pM (0.00246μg/ml). A1294 prevented IL-7 signalling through STAT5 in aconcentration-dependent manner with a median IC₅₀ of 76 pM (0.0114μg/ml). Immortalized T lymphoblast CCRF-CEM cells were stimulated with34 μg/mL (2 pM) recombinant human IL-7 (rhIL-7) in the presence orabsence of DRSPAI-L7B. STAT5 phosphorylation was assessed by MSD on celllysates. DRSPAI-L7B potently blocked IL-7-induced pSTAT5 (Table 6;IC₅₀<1 pM).

TABLE 6 Summary of potency of DRSPAI-L7B in functional assays IL-7DRSPAI-L7B IC₅₀ Assay ng/ml pM ng/ml pM pSTAT5 in CCRF-CEM cells 0.034 2<0.15 <1 pSTAT5 in whole blood 1 58.8 5.1 34 T_(eff) and T_(mem)proliferation 20 1,176 78 520 (CD4⁺)^(§) T_(eff) and T_(mem)proliferation 10 588 79.5 530 (CD4⁺)^(§) IFN-γ production by PBMC 10 58829.4 195.8 IL-17 production by T_(mem) 20 1,176 40.5 270 ^(§)Assays wereperformed using EC80 stimulation with IL-7 determined for each batch ofcells

IL-7 Induced STAT5 Phosphorylation in Healthy and Disease T Cells

PBMCs from healthy donors or IBD patients (two Crohn's disease and oneulcerative colitis) were stimulated with rhIL-7 in the presence ofDRSPAI-L7B or anti-RSV antibody (isotype control). Stimulated cells werefixed and STAT5 phosphorylation in CD8⁺ (FIG. 2B), CD4⁺ (FIG. 2C), andCD3⁺ (FIG. 2D) T cells was assessed by flow cytometry. Data is shown aspSTAT5 increase relative to unstimulated condition, mean±SD, n=3.*p<0.05, **p<0.01, ***p<0.001, matched 2-way ANOVA with Sidak's multiplecomparisons correction. FIG. 2B, FIG. 2C and FIG. 2D.

Example 4: Dynamic Light Scattering Assay

Dynamic Light Scattering (DLS) analysis was carried out on DRSPAI-L7B,A1290, A1291 and A1294 to characterize high order aggregate species andsample heterogeneity.

Each of the respective antibodies was concentrated to ≥10 mg/ml andbuffer exchanged via dialysis into 50 mM sodium phosphate pH 7.5 or 50mM sodium acetate pH 5.0 buffers followed by normalization to 10 mg/mland filtration using a 0.22 μm syringe filter. Post filtration all thesamples were stressed at a target concentration of 10 mg/ml for 2 weeksat 40° C. Samples were evaluated following incubation under stressed andunstressed conditions in the two buffers. 100 μl samples were run intriplicate at 25° C. on a Wyatt DynaPro DLS Plate reader using 96 wellCorning Costar 3635 plates sealed with Corning 6575 seals.

The data was analysed using DYNAMICS v7.1.9 software. The data wasdistributed into the following peaks: Peak 1=0.1-1 nm, Peak 2=1-10 nm,Peak 3=10-100 nm, Peak 4=100-1000 nm, Peak 5=1000-10000 nm. The data wasfiltered using the following criteria: amplitude must be between 0 and1, baseline limit 1±0.01 and sum of all squared (SOS) must be less than100. An Rh of >8 nm and average % mass at Peak2<98% was associated witha severe aggregation risk.

A1290, A1291 and A1294 showed severe aggregation risks, all exhibitinghydrodynamic radius (Rh) values of >8 nm. DRSPAI-L7B had an Rh value of<8 nm and did not, thus, exhibit such aggregation risk.

ABBREVIATIONS

γc Common gamma chain BV510 Brilliant Violet 510 CD(X) Cluster ofdifferentiation (X) CHO Chinese hamster ovary cells CST Cytometer set-upand tracking DMSO Dimethylsulphoxide F.A.C Final assay concentrationFACS Fluorescence-activated cell sorting FcR Fe receptor FCS Foetal calfserum FITC Fluorescem isothiocyanate FSC-(A/H/W) Forward scatter -(area/height/width) HEK Human embryonic kidney cells IBD Inflammatorybowel disease IC₅₀ Inhibitory concentration 50% IL-7 Interleukin 7IL-7Rα Interleukin 7 receptor alpha JAK Janus kinase (e)LNB (Electronic)laboratory notebook mAb Monoclonal antibody PBMCs Peripheral bloodmononuclear cells PBS Phosphate buffered saline PE PhycoerythrinPerCP/Cy5.5 Peridinin chlorophyll protein complex/Cyanine 5.5 SDStandard deviation SEM Standard error of the mean SSC-(A/H/W) Sidescatter - (area/height/width) pSTAT5 Phosphorylated STAT5 rhIL-7Recombinant human IL-7 RPMI Roswell Park Memorial Institute medium STAT5Signal transducer and activator of transcription 5

Example 5: Impact of DRSPAI-L7B On Cytokine Production in Human PBMC

DRSPAI-L7B was used to interrogate the role that IL-7 plays in Th1 andTh17 function and differentiation. The effect of DRSPAI-L7B on cytokinesecretion from healthy PBMCs stimulated with IL-7 in the presence of aCD3 agonistic antibody was assessed.

Healthy volunteer blood was provided by a Blood Donation Unit: Blood waswithdrawn by venepuncture and transferred into a pot or blood bagcontaining sodium heparin (1 U/mL). The blood was collected and used forPBMC isolation.

The blood was diluted 2× with PBS, layered onto 15 mL ficoll in Accuspintubes and centrifuged at 800 rcf for 20 minutes without break. Theplasma was removed carefully with a pipette and the layer containing thePBMCs was carefully transferred to a 50 mL tube. The PBMCs were washedtwice in 50 mL PBS (250 rcf, 10 minutes) and then resuspended in 50 mLRPMI+10% FCS+L-glutamine and counted using Vi-cell XR.

PBMCs were resuspended in culture media at 5×10⁶ cells/mL. Antibodydilutions of anti-RSV antibody (isotype control) and DRSPAI-L7B wereprepared at 2.5× the required final concentration in culture media.Equal volumes of cell suspension and antibody dilutions (150 μL) weremixed and incubated for 30 min at room temperature. IL-7 was prepared at5× the final concentration in culture medium and 20 μL added in therequired wells of a 96-well U bottom polystyrene plate, which had beenpre-coated with 10 μg/mL anti-CD3 at 4° C. overnight. Culture medium wasadded in all remaining wells. 80 μL of the mixed cellsuspension/antibody dilutions (2×106 cells/well) were added to each wellof the 96-well plate and left to incubate for 48 hrs at 37° C., 5% CO2.

After 48 hrs the plate was centrifuged at 300 rcf for 5 minutes and thesupernatants were removed without disturbing the pellets and transferredto a new 96-well U-bottom plate. Supernatants were either used forELISA/MSD immediately after harvesting or stored at −80° C. untilfurther use.

Anti-RSV IgG1 was at stock concentration of 2.8 mg/mL. Thawed anddiluted to the appropriate concentration on day of use. DRSPAI-L7B stockconcentration was 11.33 mg/mL. Thawed and diluted to the appropriateconcentration on day of use. Recombinant Human IL-7 was purchased fromR&D systems. The lyophilized protein was resuspended to 25 g/mL insterile PBS+0.1% bovine serum albumin and 50 μL aliquots stored at −20°C. Thawed and diluted to the appropriate concentration on day of use.Anti-CD3 clone HIT3a. Stock was 1 mg/mL. Diluted to the appropriateconcentration on day of use.

Other Materials

Reagent Company RPMI Gibco FCS In house L-glutamine In house 96-wellflat bottom type TC treated polystyrene plates Costar 96-well U bottomtype TC treated polystyrene plates Greiner DPBS Gibco Aluminium FoilLids Beckman Coulter Plate sealers Greiner bio-one U-plex BiomarkerGroup 1 (Human) Mesoscale Discovery

ECL signals were derived from the MSD instrument and converted toconcentrations using the standard curve for each analyte. For graphs inwhich the data is normalized, the average of anti-CD3⁺ IL-7 samples wasset at 100% and all other values were normalized to that. The antibodyconcentrations were log transformed and plotted against cytokineconcentrations. The no IL-7 (anti-CD3 only) samples are shown in eachgraph for comparison. For curve fitting, the following nonlinear fitfrom Graphpad Prism was used: log(inhibitor) vs. response (threeparameters) and IC₅₀ values were calculated by Graphpad Prism.

DRSPAI-L7B inhibited IFN-γ (FIG. 3A) and IL-10 (FIG. 3B) secretion in aconcentration-dependent manner (IFN-γ mean IC₅₀=195.8±101 pM; IL-10 meanIC₅₀=207.2±86 pM). Data represent the mean of n=6 independentdonors±SEM, assessed in 3 independent experiments. FIG. 3C, FIG. 3D,FIG. 3E, FIG. 3F, FIG. 3G illustrates inhibition of IL-2 by DRSPAI-L7Bin the presence of rhIL-7 and anti-CD3. There was an increase in IL-2production following IL-7 stimulation in 5 donors. This increase wasinhibited fully by DRSPAI-L7B.

Example 6: DRSPAI-L7B Inhibits Cytokine Production by Memory T Cells

To accurately determine the role of IL-7 in Th17 cell function anddifferentiation, a ‘poised Th17’ assay was used to profile the secretionof Th17-associated cytokines. T_(mem) cells were isolated from healthydonors and incubated with IL-7 in the presence of DRSPAI-L7B.

Whole human blood was collected from donors in the Blood Donation Unit(BDU). Typically, 200 mL was collected by venepuncture per donor and ananti-coagulant, citrate-dextrose solution (ACD, Sigma, Cat #C3821) wasadded immediately to each sample. ACD was added at 15% (e.g. 30 mL ACDadded to 200 mL blood). Within 2 hours of collection, and using amicrobiological safety cabinet, the blood was dispensed evenly intopre-filled LEUCOSEP tubes (Greiner, Cat #227288) at 30 mL (max) pertube. The blood was centrifuged for 15 minutes at 800×g at roomtemperature in a swing bucket rotor with no brake applied. The PBMCswere washed in PBS (500×g, 10 minutes) and then resuspended in 10 mL PBSand counted using the NucleoCounter. The cells were centrifuged again at500×g and the cell pellet was re-suspended in FACS buffer at aconcentration of 5×10∂cells/mL.

Human CD4⁺ memory T cells (T_(mem)) EasySep enrichment cocktail wasadded at 50 μL/mL to the cell suspension. The cells were incubated atroom temperature for 10 minutes after which the EasySep magneticparticles were added at 50 μL/mL. The suspension was again incubated atroom temperature for 10 minutes after which the cell suspension wasplaced in an EasySep magnet for 10 minutes. The cell suspension was thentransferred to a fresh tube whilst still placed within the magnet toensure a negative selection. The cells were centrifuged at 500×g for 10minutes and resuspended in cell culture media. The cell count wasdetermined using the NucleoCounter.

The cells were rested at 37° C. and 5% CO₂ overnight in culture media.The next day the cells were centrifuged and resuspended in assay mediaat a concentration of 2.5×10{circumflex over ( )}5 cells/mL. RecombinanthIL-7 was added to the cells at a final concentration of 20 ng/mL. Thecells were then incubated in the presence of DRSPAI-L7B for 4 days at37° C. and 5% CO₂ after which they were stimulated with 10 nM PMA and 1M lonomycin for 16 hours at 37° C. and 5% CO₂.

Detection of IL-17 from the Supernatants Using MSD

After 16 hours of PMA and ionomycin stimulation, the cells werecentrifuged and 40 μL of the supernatants were transferred to the MSDplates which were pre-blocked with 0.5% of Blocker B. These MSD plateswere coated with an anti-IL-17A capture antibody. The samples wereincubated for 2 hours at room temperature whilst shaking after which theplates were washed with PBS and 0.05% Tween-20. 10 μL of the IL-17specific detection antibody labelled with the MSD SULFO-TAG reagent wasthen added and the samples were incubated for a further 2 hours at roomtemperature whilst shaking. The plate was washed again three times inPBS and 0.05% Tween-20 and 2× Read Buffer T was added to the samples.The plate was read on the Sector Imager.

Detection of Other Cytokines

A U-Plex MSD Kit was designed to enable investigation of the effect ofDRSPAI-L7B on the secretion of IL-6, IL-10, IFN-γ, TNF-α and CCL3. TheMSD U-Plex plates were prepared by coating the plate with linker coupledcapture antibodies. Each capture antibody was biotinylated and had aunique linker assigned to it. The supernatants were diluted 1 in 100 andtransferred to the MSD plates. The samples were incubated for 1 hour atroom temperature whilst shaking after which the plates were washed withPBS and 0.05% Tween-20. 50 μL of the specific detection antibodieslabelled with the MSD SULFO-TAG reagent was then added and the sampleswere incubated for a further hour at room temperature whilst shaking.The plate was washed again three times in PBS and 0.05% Tween-20 and 2×Read Buffer T was added to the samples. The plate was read on the SectorImager.

DRSPAI-L7B antibody was produced at a stock concentration of 11.33 mg/mLin 20 mM Histidine, 180 mM Trehalose, 40 mM Arginine, 8 mM Methionine,0.05 mM EDTA, pH 6.0. The Antibody was diluted to a concentration of 20μg/mL in assay media. A 1 in 3 serial dilution was then carried out inassay media to generate a 10 point dose response curve. The dilutionswere transferred to the assay plates ensuring that the highest finalassay concentration of the antibody was 10 μg/mL.

BRL-54319MM (Rapamycin) was used as the positive control in the assay at1 pM final assay concentration in assay media.

Recombinant Human IL-7 was resuspended to 25 μg/mL in sterile PBS and 50μL aliquots stored at −20° C. The aliquots were thawed and diluted tothe appropriate concentration on day of use.

Other Materials

Reagent Company Pre-Filled Leucosep Tubes Greiner Human IL-17 Base KitMSD Easy50 EasySep Magnet Stemcell Phospho buffered saline, no Ca2+/Mg2+(PBS) Gibco IMDM Gibco 96 well Costar round bottom polystyrene, sterileCostar plates, with lid, clear Heat Inactivated Fetal Bovine SerumHyclone Xvivo15 Lonza Penstrep Invitrogen Easysep Human Memory CD4+T-cell enrichment Kit Stem cell L-Glutamine Invitrogen MEM Non-EssentialAmino Acids Invitrogen HEPES Invitrogen PMA-Phorbol 12-myristate13-acetate Sigma Ionomycin Sigma Sodium Pyruvate Invitrogen U-plexBiomarker Group 1 (Human) Kit Mesoscale Discovery CellTiter-Glo Promega

FACS Buffer: sterile PBS containing 2% heat-inactivated FBS. CellCulture Media: 450 mL IMDM, 50 mL FBS, 5 mL Penstrep, 5 mL L-Glutamine,5 mL Non-essential Amino Acids and 5 mL Sodium Pyruvate. Assay Media:500 mL Xvivo 15, 5 mL Penstrep, 5 mL L-glutamine, 5 mL HEPES and 5 mLSodium Pyruvate.

All data was normalized to the mean of 8 high and 8 low control wells oneach plate. A four-parameter curve fit of the following form was thenapplied.

$y = {\frac{a - d}{1 + \left( {{x/c}\text{?}} \right.} + d}$?indicates text missing or illegible when filed

Where a is the minimum, b is the Hill slope, c is the XC50 and d is themaximum. Data was presented as the mean IC₅₀ with the standard deviationof the mean of n experiments.

CD4⁺ T_(mem) cells, isolated from healthy donor blood, were incubatedwith IL-7 in the presence of DRSPAI-L7B for 4 days after which they werespiked with PMA/ionomycin for 16 hours before harvest.

Cytokine secretion into the supernatant was assessed by MSD. DRSPAI-L7Btreatment resulted in a concentration-dependent inhibition of IL-17(FIG. 4A, IC₅₀=270±31.59 pM), TNFα (FIG. 4B, IC₅₀=127.1±89.83 pM), IL-6(FIG. 4C, IC₅₀=202.07±99.64 pM), IL-10 (FIG. 4D, IC₅₀=197.48±148.91),INFγ (FIG. 4E, IC₅₀=157.98±89.55) and CCL3 (FIG. 4F, IC₅₀=163.83±85.96)secretion. Dose dependent inhibition was also observed for IL-6, IL-10,IFNγ, TNFα and CCL3. Inhibition of the production of these cytokines wasexpressed as a percentage of that achieved with Rapamycin (1 pM)positive control.

Example 7: T Cell Population Analysis

Given the central role of T cells in MS, T cell populations wereanalysed in PBMCs from RRMS, PPMS and SPMS patients.

All human samples were obtained with patient informed consent inaccordance with ICH GCP under a protocol approved by a national,regional or investigational center ethics committee or an InstitutionalReview Board (IRB) approved protocol.

Healthy PBMCs were isolated from BDU blood and stored frozen in liquidnitrogen until used. Disease PBMCs were supplied by an approved externalhuman tissue supplier. Healthy control blood was withdrawn byvenepuncture and transferred into a container with sodium heparinanti-coagulant (1 U/mL). The blood was collected and used within 1 hourfor PBMC isolation.

Healthy Control human PBMCs prepared and frozen in advance were usedPBMCs were prepared by layering blood on 15 mL Ficoll. Tubes werecentrifuged at 800 g for 20 minutes, with brake off. The mononuclearcell layer at the interface was transferred to 50 mL Falcon tubes,washed by topping up to 45 mL with PBS and centrifuging at 300 g for 10minutes. The pellets were resuspended in Freezing medium A (60:40FCS:medium), 5% of the original blood volume and then an equal volume ofFreezing medium B (80:20 FCS:DMSO) was added, dropwise to reduce osmoticshock. Cells were transferred to cryovials (1 mL per vial (around 1×10⁷cells)) and frozen in a Mr Frosty freezing container at −80° C. for upto 1 week, followed by transfer to liquid nitrogen for long termstorage.

PBMC Recovery

Cells were thawed by removing from liquid nitrogen storage andimmediately placed in a water bath at 37° C. until thawed. Aftertransferring cell suspension to a 50 mL centrifuge tube, medium(RPMI+10% heat-inactivated FCS, 1% penicillin/streptomycin, and 1%glutamine) was added very slowly to decrease the DMSO concentrationgradually. Once the volume was increased to 30 mL the cells werecentrifuged, 300 g for 10 minutes, and resuspended in 5 mL medium beforecounting, made up to 15 mL with medium, centrifuged as above and cellsresuspended in an appropriate volume of medium to yield 5×10⁶ cells/1mL.

In each of 5 individual experiments, two healthy control (HC), two RRMS,one PPMS and one SPMS, donor PBMC sample were thawed as above and cellstreated as below. Different donors were used for each experiment, HCsbroadly age and gender matched with the disease patient donors used.

T Cell Phenotyping Flow Cytometry Assay

Following resuspension of PBMCs at 5×10⁶/mL in medium, 100 μL cells weretransferred to FACS tubes (for full stain) and an additional 50 μL of HCsamples (for FMO control tubes). Cells were washed by addition of 2 mLFACS buffer, centrifuged 300 g, 5 minutes and pellet resuspended inresidual volume. 5 μL human FcX Trustain block was added for 10 minsfollowed by addition of 100 μL antibody stain cocktail and incubated atroom temperature for 30 mins. Cells were washed by addition of 2 mL FACSbuffer, centrifuged 300 g, 5 minutes and pellet resuspended in residualvolume. 500 μL diluted Live/Dead Fixable Aqua Dead Cell Stain was addedand incubated at room temperature for 25 mins. Cells were washed byaddition of 2 mL FACS buffer, centrifuged 300 g, 5 minutes and pelletresuspended in residual volume. 200 μL FACS buffer was added and samplesanalyzed on the same day, using a BD FACS Canto II.

The performance of the instrument was checked using the Cytometer Set-upand Tracking (CST) Beads. This is a QC check for the instrument, setsthe baselines and optimizes the voltages for each laser prior to use.The results of the calibration are stored within the CST software on theinstrument.

Compensation for the instrument was performed using UltraCompcompensation beads in accordance with the manufacturer's instructions.The relevant antibodies used to stain the cells during the experimentwere used to label the compensation beads. Compensation for theexperiments were performed using the appropriately labelled beads withthe automatic compensation facility available within the FACS Divasoftware. After analysis of the compensation samples the compensationsettings were calculated and applied to each experimental stainingpanel.

Reagent PBS (w/o Ca²⁺ and Mg²⁺) RPMI 1640 L-Glutamine 200 mM Pen/StrepHeat inactivated FCS DMSO FcX Trustain blocking reagent BD HorizonBrilliant Stain Buffer anti-human CD8 AF488 (clone; RPA-T8) anti-humanCD25 PE (clone: BC96) anti-human CD4 PerCP/Cy5.5 (clone: RPA-T4)anti-CD4SRO PE/Cy7 (clone: UCHL1) anti-CCR7 AF647 (clone: G043H7)anti-CD20 APC/Fire750 (clone: 2H7) anti-CD127 BV421 (clone: A019D5)anti-CD14 BV510 (clone: M5E2) anti-CD19 BV510 (clone: SJ25C1) anti-CD56BV510 (clone: 5.1H11) anti-CD16 BV510 (clone: 3G8) Aqua fixableLive/Dead stain UltraComp eBeads Compensation Particles Set BD FACSDivaCS&T Research Beads

Equipment Description Muse Cell Analyser (counter) BD Canto II FlowCytometer

Samples were acquired on a BD FACS Canto II flow cytometer using BDBioSciences FACS Diva software (v8.0.1). Resulting compensated .fcsfiles were analyzed with FlowJo software (v10.0.8) and results generatedin Excel using the Batch Analysis facility within the software.

T cell populations were analyzed in PBMCs from RRMS, PPMS and SPMSpatients. Data was generated from patients where all patients were ontreatment (10/10 RRMS patients on natalizumab and all progressive MSpatients were on steroids and/or symptomatic treatments), a reduction inT_(reg) cells was observed. CD4⁺ (FIG. 5A), CD8⁺ (FIG. 5B) andregulatory T cells (FIG. 5C) from healthy controls and MS patients wereprofiled by flow cytometry based on CD45RO, CCR7, CD127 and CD25expression on the cell surface. No difference was seen between healthyand disease T cell populations in either CD4⁺ or CD8⁺ subsets. The RRMSand PPMS T_(reg) population was significantly reduced compared to HC, nodifference was found in T_(reg) numbers in SPMS patients. ***0<0.0001,*0<0.05 as tested by One-way ANOVA with Dunnett multiple comparisontest. Data presented represent the mean±SEM of n=5-10 donors per group,analyzed in 5 independent experiments. Effector memory=CD45RO⁺CCR7⁻,central memory=CD45RO⁺CCR7⁺, naïve=CD45RO⁻CCR7⁺, effector=CD45RO⁻CCR7⁻,T_(reg)=CD127^(low/−)CD25⁺.

Example 8: STAT5 Phosphorylation

All human samples were obtained with patient informed consent inaccordance with ICH GCP under a protocol approved by a national,regional or investigational center ethics committee or an InstitutionalReview Board (IRB) approved protocol.

Healthy PBMCs were isolated from BDU blood and stored frozen in liquidnitrogen until used. Disease PBMCs were supplied by an approved externalhuman tissue supplier.

Healthy control blood was withdrawn by venepuncture and transferred intoa container with sodium heparin anti-coagulant (1 U/mL). The blood wascollected and used within 1 hour for PBMC isolation.

Healthy Control human PBMCs prepared and frozen in advance were usedPBMCs were prepared by layering blood on 15 mL Ficoll. Tubes werecentrifuged at 800 g for 20 minutes, with brake off. The mononuclearcell layer at the interface was transferred to 50 mL Falcon tubes,washed by topping up to 45 mL with PBS and centrifuging at 300 g for 10minutes. The pellets were resuspended in Freezing medium A (60:40FCS:medium), 5% of the original blood volume and then an equal volume ofFreezing medium B (80:20 FCS:DMSO) was added, dropwise to reduce osmoticshock. Cells were transferred to cryovials (1 mL per vial (around 1×10⁷cells)) and frozen in a Mr Frosty freezing container at −80° C. for upto 1 week, followed by transfer to liquid nitrogen for long termstorage.

Cells were thawed by removing from liquid nitrogen storage andimmediately placed in a water bath at 37° C. until thawed. Aftertransferring cell suspension to a 50 mL centrifuge tube, medium(RPMI+10% heat-inactivated FCS, 1% penicillin/streptomycin, and 1%glutamine) was added very slowly to decrease DMSO concentrationgradually. Once the volume was increased to 30 mL, the cells werecentrifuged (300 g for 10 minutes) resuspended in 5 mL medium andcounted, then topped up to 15 mL with medium, centrifuged as above andcells resuspended in an appropriate volume of medium to yield 5×10⁶cells/mL.

In each of 5 individual experiments, two healthy control (HC), two RRMS,one PPMS and one SPMS donor PBMC sample were thawed as above and cellstreated as below. Different HC donors were used for each experiment,broadly age and gender matched with the disease patient donors used.

Inhibition of IL-7 Induced pSTAT5 by DRSPAI-L7B in T Cells

Following resuspension of PBMCs at 5×10⁶/mL in medium, 450 μL cells weretransferred to 15 mL Falcon tubes and incubated with 5 mL PBS containing5 μL Near InfraRed live/dead stain. Cells were washed by the addition of9 mL full culture medium, centrifuged (300 g, 5 minutes) and the cellpellet resuspended in AIM V serum free culture medium at 5×10⁶ cells/mL.All antibody treatments and rhIL-7 stimulations were made up at 4× finalassay concentration in AIM V serum free culture medium before mixing1:1, (IL-7:mAb) and incubating at room temperature for 10 minutes. Inthe absence of either antibody or IL-7 stimulus, culture medium wasadded. Final concentration of IL-7 used was 1 ng/mL (57 pM). Finalconcentration of mAb used: 500 ng/mL (3.33 nM).

100 μL of PBMC suspension was added to four FACS tubes per donor (5×105cells/test). 100 μL of the antibody:IL-7 mixture was added to theappropriate FACS tubes (medium alone, IL-7 alone, IL-7+DRSPAI-L7B orIL-7+anti-RSV isotype control antibody). Tubes were mixed gently beforeincubating at 37° C. in a humidified incubator for 20 minutes. At theend of the stimulation period, the cells were centrifuged at 300 g for 5minutes, the cell pellet resuspended and 250 μL of pre-warmed PHOSFLOWfixation buffer (1×) added. The samples were incubated for a further 10minutes at 37° C. Following fixation, cells were pelleted bycentrifugation (300 g, 5 minutes) and washed in 2 mL PBS. Cell pelletswere resuspended in 100 μL ice cold Perm Buffer III and gently vortexingto mix. Cells were incubated on ice for 30 minutes before washing oncein 1 mL PBS and centrifuging (300 g, 5 minutes). Cells were washed in 2mL PBS then centrifuged (300 g, 5 minutes).

Following permeabilization cell pellets were resuspended in 25 μL FcRblocking reagent diluted 1:5 in FACS buffer. Cells were incubated for 10minutes at room temperature before adding the detection antibodystaining cocktail: 5 μL anti-CD3 (PerCP/Cy5.5), 5 μL anti-CD4 (AF488), 5μL anti-CD8 (APC), 20 μL anti-pSTAT5 (PE) and 15 μL FACS buffer (50 μLtotal per test). Where a stain was excluded for FMO controls, theequivalent volume of FACS buffer was added instead. Tubes were mixedbriefly and incubated at room temperature (RT) for 30 minutes, protectedfrom light. Cells were washed in 2 mL FACS buffer, centrifuged (300 g, 5mins) and pellets resuspended in 100 μL FACS buffer for analysis on thesame day using a FACS Canto II flow cytometer.

The performance of the FACS Canto II instrument was checked using theCytometer Set-up and Tracking (CST) Beads. This is a QC check for theinstrument, sets the baselines and optimizes the voltages for each laserprior to use. The results of the calibration are stored within the CSTsoftware on the instrument.

Compensation for the instrument was performed using UltraCompcompensation beads in accordance with the manufacturer's instructions.The relevant antibodies used to stain the cells during the experimentwere used to label the appropriate compensation bead type. Forcompensation of live/dead cell dye, 1 μL of undiluted dye was added to 1drop of ArcAmine reactive beads, incubated for 30 minutes, washed andArcAmine negative beads added immediately before running sample.Compensation for the experiments were performed using the appropriatelylabelled beads with the automatic compensation facility available withinthe FACS Diva software. After analysis of the compensation samples thecompensation settings were calculated and applied to each experimentalstaining panel.

Antibody and Reagents

For experimentation, antibody aliquots were thawed and stored at WC forno longer than 8 weeks. DRSPAI-L7B (anti-IL-7) and anti-RSV Isotypecontrol.

Reagent Supplier PBS (w/o Ca²⁺ and Mg²⁺) Life Technologies RPMI 1640Life Technologies L-Glutamine 200 mM Life Technologies Pen/Strep LifeTechnologies Heat inactivated FCS Life Technologies AIM V Medium GibcoRecombinant human IL-7 R&D Systems BD Phosflow ™ Lyse/Fix Buffer 5x BDBioscience BD Phosflow ™ Perm Buffer III BD Biosciences FcX Trustainblocking reagent BioLegend BD Phosflow ™ PE Mouse anti-STAT5 (pY694) BDBioscience Mouse anti-human CD8 APC (clone: SK1) BioLegend Mouseanti-human CD4 AF488 (clone: RPA-T4) BioLegend Mouse anti-human CD3PerCP/Cy5.5 (clone: SK7) BioLegend Near Infra-Red fixable Live/Deadstain Life Technologies UltraComp eBeads Compensation Particles Set LifeTechnologies ArcAmine reactive beads Life Technologies BD FACSDiva CS&TResearch Beads BD Biosciences

Equipment Description Muse Cell Analyser (counter) BD Canto II FlowCytometer

Samples were acquired on a BD FACS Canto II flow cytometer using BDBioSciences FACS Diva software (v8.0.1). Resulting compensated .fcsfiles were analyzed with FlowJo software (v10.0.8) and results generatedin Excel using the Batch Analysis facility within the software.

In order to determine the capacity for disease cells to respond to IL-7and confirm the efficacy of DRSPAI-L7B in samples from MS patients,PBMCs isolated from either healthy controls or MS patient donors werestimulated with 1 ng/mL (58 pM) IL-7 in the presence of a non-saturating(IC90=500 ng/mL (3.3 nM)) concentration of DRSPAI-L7B or anti-RSV IgG1κisotype control antibody.

PBMCs from healthy donors or MS patients (10 Healthy, 10 RRMS, 5 PPMSand 5 SPMS) were stimulated with rhIL-7 in the presence of DRSPAI-L7B oranti-RSV antibody (isotype control). STAT5 phosphorylation in CD4⁺ (FIG.6A) and CD8⁺ (FIG. 6B) T cells was assessed by flow cytometry. IL-7stimulation induced STAT5 phosphorylation in CD4⁺ and CD8⁺ T cellsderived from healthy donors and MS patients with a 1 ng/mL (58 pM,˜200-fold higher than reported in disease). IL-7 stimulus was almostentirely abrogated by 500 ng/mL or 3.3 nM (non-saturating concentration)of DRSPAI-L7B. Data shown is normalized to IL-7 treatment condition,mean±SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. One-way ANOVAwith Tukey's multiple comparison correction. Similar results wereobserved upon IL-7 stimulation of PBMCs derived from IBD patients.

Example 9: IL-7 Levels in Disease

All human samples were obtained with patient informed consent inaccordance with ICH GCP under a protocol approved by a national,regional or investigational center ethics committee or an InstitutionalReview Board (IRB) approved protocol. Samples were stored at −80° C.until required for use.

The measurement of IL-7 was conducted following the method SOP. Briefly,the method follows the MSD kit protocol with a sample dilution of 1:2with Diluent 43. Diluted samples were incubated on a pre-coated IL-7 MSDplate followed by detection with a ruthenylated anti-IL-7 antibody. Thelevel of IL-7 in the plasma sample is directly proportional to theresulting ECL signal read using the MSD Sector Imager 6000 Reader. TheIL-7 method uses the MSD V-PLEX human IL-7 kit (catalogue no. K151RCD).

ECL signals for standards, controls, and unknown samples were derivedfrom the MSD instrument and exported to Softmax Pro GxP for analysis.The back-calculated concentrations for each method were interpolatedusing the standard curve and a 4-parameter curve fitting model with 1/γ2weighting. A summary of the IL-7 concentrations was transferred toMicrosoft Excel.

All data were transferred from Microsoft Excel 2016 to GraphPad Prism v.6 for graphing and statistical analysis. Data were copied into “ColumnTables” in Prism based on sample matrix (serum) and analyte (IL-7).Column statistics tests were carried out on each set of data includingShapiro-Wilk normality test to determine whether each data set wasnormally distributed. If data were not normally distributed, data weretransformed using the formula γ=log(γ) to normalize distribution. Columnstatistics testing was carried out again on the transformed data inorder to confirm normal distribution.

For serum samples, groups were compared using one-way analysis ofvariance (ANOVA). Distribution of the data was tested using theShapiro-Wilk normality test in the column statistics function. Groupdifferences in non-normally distributed data were assessed usingKruskal-Wallis unpaired, non-parametric analysis with Dunn's multiplecomparisons test to compare the mean rank of each column with oneanother. Data were also transformed using γ=log(γ) in order to normalizethe distribution of the data. Following transformation, Shapiro-Wilktest was carried out using the column statistics function to confirmthat transformation resulted in normal distribution of the data.Transformed, normally distributed data was then assessed forstatistically significant differences using an ordinary one-way,unpaired ANOVA with Tukey's multiple comparison test to compare the meanof each column to one another.

IL-7 levels were quantified in serum samples from healthy controls (HC,n=10), Crohn's disease (n=15), ulcerative colitis (UC, n=15), systemiclupus erythematosus (SLE, n=15) and primary Sjögren's syndrome (pSS,n=15) patients. IL-7 was significantly increased in Crohn's disease, UCand SLE (7.3, 9.44 and 5.53-fold, respectively) compared to healthycontrols. There was also a small increase in IL-7 in the pSS cohort(3.55-fold vs HC). Data are mean±SD. **p≤0.01, ***p≤0.001, ****p≤0.0001;Serum analyzed using 1-way ANOVA with Dunn's multiple comparisonscorrection. FIG. 7A and FIG. 7B.

Example 10: DRSPAI-L7B Production

Cell lines were transfected with a plasmid. This plasmid contained thecodon optimized DRSPAI-L7B heavy and light chain genes, each under thetranscriptional control of separate human EF1α promoters. The lightchain constant region is human kappa and the heavy chain constant regionis human IgG1, containing the LAGA substitution. LAGA substitutioncorresponds to L235A/G237A. Transfected cell lines were expanded andtriaged based on expression of DRSPAI-L7B.

Example 11: Epitope Binding Instrumentation

-   -   Waters Synapt G2-Si mass spectrometer    -   Acquity M Class UPLC    -   LEAP H/D-X PAL liquid-handling robot

Solutions

-   -   Quench solution: 400 mM potassium phosphate, 6 M guanidine        hydrochloride, 0.5 M TCEP pH 2.5 (after 1:1 mixing with        sample—quench buffer pH-adjusted with NaOH to give this pH on        mixing).    -   Dilution buffer:50 mM Na phosphate 100 mM NaCl in H2O pH7.0.

Proteins:

-   -   IL7: Concentration: 0.68 mg/ml (33 pM) in PBS. Sequence before        processing (SEQ ID NO:48):

MFHVSFRYIFGLPPLILVLLPVASSDCDIEGKDGKQYESVLMVSIDQLLDSMKEIGSNCLNNEFNFFKRHICDANKEGMFLFRAARKLRQFLKMNSTGDFDLHLLKVSEGTTILLNCTGQVKGRKPAALGEAQPTKSLEENKSLKEQKKLNDLCFLKRLLQEIKTCWNKILMGTKEHHHHHH

-   -   Secretory leader (underlined above)    -   DRSPAI-L7B was produced in HEK cells and used at 15 mg/ml (100        μM) in PBS.

HDX

-   -   Dilutions buffers were:        -   Non-deuterated: 50 mM Na phosphate 100 mM NaCl in H2O pH7.0        -   Deuterated: 50 mM Na phosphate 100 mM NaCl in D2O pD 6.6    -   For initial testing, DRSPAI-L7B was tested in a single run to        check for digestion quality and signal strength.

For the HDX experiment, IL7:mAb mixture was prepared as follows:

“Apo” sample comprised 20 μl concentrated IL7, 15 μl PBS and 25 μldilution buffer. “mAb” samples comprised 20 μl concentrated IL7, 15 μlDRSPAI-L7B and 25 μl dilution buffer. This gave final nominalconcentrations of 50 μM for IL7 and 25 μM for DRSPAI-L7B. Samples wereprepared on ice and kept at 0° C. until analysis.

For the mAb binding experiments, samples were subjected to a standarddeuteration method using 10-fold dilution into deuteration buffer,performed using the LEAP H/D-X PAL robot. Protein samples were placed inthe 0° C. rack. 6 μl protein sample was transferred to a vial held at20° C., then 54 μl dilution buffer (non-deuterated for 0 time point,deuterated for other time points) was added to initiate hydrogenexchange. After incubation for the appropriate time, 50 μl of thissample was aspirated and transferred to a pre-cooled vial (0° C.)containing 50 μl quench solution (400 mM sodium phosphate, 6 M guanidinehydrochloride, 0.5 M tris-carboxyethyl phosphine[pH 2.5 after mixingwith sample]). After mixing and incubation for 1 min, 90 μl of thissample was re-aspirated and transferred to the HDX manager, wherethrough a 100 μl loop it was injected onto an immobilised pepsin column(Enzymate BEH, 2.1×30 mm, Waters #186007233, held at 15° C.), at 50μl/min. Resulting peptides were eluted in 0.2% formic acid and trappedon a Vanguard BEH C18 pre-column (2.1×5 mm; Waters #186003975). The trapcolumn was then switched in line with an analytical reverse-phase column(BEH C18, 1×100 mM, Waters #186002346) and peptides eluted using thefollowing gradient, where A=0.2% formic acid, 0.03% trifluoroacetic acidin water, B=0.2% formic acid in acetonitrile:

Step Time(min) Flow rate(uL/min) % A % B Curve 1. Initial 40.000 88.012.0 2. 8.00 40.000 64.0 36.0 6 3. 9.00 40.000 5.0 95.0 6 4. 10.0040.000 5.0 95.0 6 5. 10.50 40.000 88.0 12.0 6 6. 11.50 40.000 5.0 95.0 67. 13.00 40.000 5.0 95.0 6 8. 13.50 40.000 88.0 12.0 6 9. 15.00 40.00088.0 12.0 6

During the gradient phase, the pepsin column was washed at 25 μl/minwith pepsin wash buffer (2 M guanidine-HCl, 0.8% formic acid, 5%acetonitrile 5% propan-2-ol pH2.5), 2×80 μl, and returned to 0.2% formicacid at 25 μl/min in preparation for the next sample. Eluate from theanalytical column was analyzed using ESI-MS, using a Waters Synapt G2-Simass spectrometer operating in positive, resolution mode, with continuumdata collected. Lockspray data containing Leucine-enkephalin and Glu-Fibions was also acquired. For peptide identification samples, MSe datawere acquired (acquisitions alternating between low and high energyconditions in the collision cell) to provide fragmentation data to aidrobust peptide identification. For HDX samples, a single low energyacquisition (plus lockspray) was acquired.

An initial sample was run in MSe mode to generate a peptide search listusing ProteinLynx Global Serverv3.0.2 (Waters). HDX samples were run induplicate with deuteration periods of 0, 0.5 and 5 min. The peptidesearch list was imported into DynamX v3.0 (Waters) and filtered to givehigh-quality peptides to search for in HDX samples (minimumintensity>10,000; peptide score >7.0. The HDX sample data were thenbrought in and processed to determine deuteration for each identifiedpeptide in each sample. Peptide and ion assignments were manuallychecked and refined where necessary.

To interpolate the peptide-level data to residue-level data (to enableheat maps and structural views) the algorithm used by DyanamX togenerate heat maps was used, where for each residue data from theshortest overlapping peptide was used (where two overlapping peptideswere of the same length, the peptide nearest the N-terminus was used).

Results

Two peptides in the region of residues 67-81 (numbering fromfull-length, unprocessed construct) showed strong protection in thepresence of mAb.

(SEQ ID NO: 12-FKRHICDANKEGMFL) (SEQ ID NO: 16-FKRHICDANKEGMF)

Again, the region covering the residues 67-81 showed a clear protectionsignal. Further, the heat-map data was mapped onto an available 3Dstructure for IL7 (in complex with IL7Rα), and the protected regioncovering residues 67-81 (FKRHICDANKEGMFL (SEQ ID NO: 12)). This regionwas distal from the region of IL7 interacting with the receptor IL7Rα.The epitope sits adjacent to the IL7Rα and γ-chain interaction sites onthe folded protein.

Embodiments

Other aspects and embodiments of the disclosure will be apparent fromthe exemplary embodiments that follows.

Embodiments

-   -   1. An IL-7 binding protein that binds to one or more amino acid        residue within the amino acid sequence set forth in SEQ ID NO:12        of human IL-7.    -   2. The IL-7 binding protein of embodiment 1, which protects        residues 67 to 81 (SEQ ID NO:12) of IL-7 from deuterium exchange        in HDX-MS analysis.    -   3. The IL-7 binding protein of embodiment 1, which protects        residues 67 to 80 (SEQ ID NO:16) of IL-7 from deuterium exchange        in HDX-MS analysis.    -   4. An IL-7 binding protein or an IL-7 binding fragment thereof        that binds to human IL-7 adjacent an IL-7Rα binding site, with a        KD of 100 nM or less and inhibits IL-7 binding to IL-7R as        measured in an in vitro competitive binding assay.    -   5. The IL-7 binding protein of any one of embodiments 1-4,        wherein the IL-7 binding protein comprises at least one of (a) a        heavy chain CDR1 having at least 80% identity to the amino acid        sequence set out in SEQ ID NO:6, (b) a heavy chain CDR2 having        at least 80% identity to the amino acid sequence set out in SEQ        ID NO:7 or (c) a heavy chain CDR3 having at least 80% identity        to the amino acid sequence set out in SEQ ID NO:8.    -   6. The IL-7 binding protein according to any one of embodiments        1-5, wherein the IL-7 binding protein comprises at least one        of (a) a light chain CDR1 having at least 80% identity to the        amino acid sequence set out in SEQ ID NO:9, (b) a light chain        CDR2 having at least 80% identity to the amino acid sequence set        out in SEQ ID NO:10 or (c) a light chain CDR3 having at least        80% identity to the amino acid sequence set out in SEQ ID NO:11.    -   7. The IL-7 binding protein according to any one of embodiments        1-6, wherein the IL-7 binding protein comprises a variable        region light chain having at least 80% identity to the amino        acid sequence set out in SEQ ID NO:5.    -   8. An IL-7 binding protein or an IL-7 binding fragment thereof        comprising a variable region light chain having at least 95%        identity to the amino acid sequence set out in SEQ ID NO:5.    -   9. The IL-7 binding protein according to embodiment 8, wherein        the IL-7 binding protein comprises at least one of (a) a heavy        chain CDR1 having at least 80% identity to the amino acid        sequence set out in SEQ ID NO:6, (b) a heavy chain CDR2 having        at least 80% identity to the amino acid sequence set out in SEQ        ID NO:7 or (c) a heavy chain CDR3 having at least 80% identity        to the amino acid sequence set out in SEQ ID NO:8.    -   10. An IL-7 binding protein or an IL-7 binding fragment thereof        comprising a heavy chain CDR1 having at least 80% identity to        the amino acid sequence set out in SEQ ID NO:6, a heavy chain        CDR2 having at least 80% identity to the amino acid sequence set        out in SEQ ID NO:7 and a heavy chain CDR3 having at least 80%        identity to the amino acid sequence set out in SEQ ID NO:8.    -   11. An IL-7 binding protein or an IL-7 binding fragment thereof        comprising a light chain CDR1 having at least 80% identity to        the amino acid sequence set out in SEQ ID NO:9, a light chain        CDR2 having at least 80% identity to the amino acid sequence set        out in SEQ ID NO:10 and a light chain CDR3 having at least 80%        identity to the amino acid sequence set out in SEQ ID NO:11.    -   12. The IL-7 binding protein according to any one of embodiments        10-11, wherein the IL-7 binding protein comprises a variable        region light chain having at least 80% identity to the amino        acid sequence set out in SEQ ID NO:5.    -   13. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein        comprises a variable region light chain having the amino acid        sequence set out in SEQ ID NO:5.    -   14. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein        comprises a heavy chain CDR1 comprising the amino acid sequence        set out in SEQ ID NO:6, a heavy chain CDR2 comprising the amino        acid sequence set out in SEQ ID NO:7 and a heavy chain CDR3        comprising the amino acid sequence set out in SEQ ID NO:8.    -   15. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein        comprises a light chain CDR1 comprising the amino acid sequence        set out in SEQ ID NO:9, a light chain CDR2 comprising the amino        acid sequence set out in SEQ ID NO:10 and a light chain CDR3 the        amino acid sequence set out in SEQ ID NO:11.    -   16. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein        comprises a variable region heavy chain having at least 80%        identity to the amino acid sequence set out in SEQ ID NO:4.    -   17. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein        comprises a variable region heavy chain comprising the amino        acid sequence set out in SEQ ID NO:4.    -   18. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein        comprises a light chain having at least 80% identity to the        amino acid set sequence out in SEQ ID NO:3.    -   19. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein        comprises a light chain comprising the amino acid sequence set        out in SEQ ID NO:3.    -   20. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein        comprises a constant region such that the IL-7 binding protein        has reduced ADCC and/or complement activation or effector        functionality.    -   21. The IL-7 binding protein according to embodiment 20, wherein        the IL-7 binding protein comprises a heavy chain Fc domain        having an alanine residue at position 235 and position 237        according to EU numbering.    -   22. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein        comprises a heavy chain having at least 80% identity to the        amino acid sequence set out in SEQ ID NO:2.    -   23. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein        comprises a heavy chain comprising the amino acid sequence set        out in SEQ ID NO:2.    -   24. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein        comprises a heavy chain comprising the amino acid sequence set        out in SEQ ID NO:2 and a light chain comprising the amino acid        sequence set out in SEQ ID NO:3.    -   25. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein is an        antibody or an antigen binding portion thereof.    -   26. The IL-7 binding protein according to embodiment 25, wherein        the antibody is a monoclonal antibody.    -   27. The IL-7 binding protein according to embodiment 26, wherein        the monoclonal antibody is an IgG1.    -   28. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein is        human, humanized or chimeric.    -   29. The IL-7 binding protein according to embodiment 28, wherein        the IL-7 binding protein is humanized.    -   30. The IL-7 binding protein according to embodiment 28, wherein        the IL-7 binding protein is human.    -   31. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein binds to        and neutralizes IL-7.    -   32. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein binds to        native IL-7.    -   33. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein binds to        circulating IL-7.    -   34. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein is an        isolated IL-7 binding protein.    -   35. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein binds to        IL-7 and inhibits IL-7 depending IFN-γ or IL-10 secretion from        peripheral blood mononuclear cells with an IC50 of 1 nM or less.    -   36. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein binds to        IL-7 and inhibits IL-7 depending STAT5 phosphorylation in CD4+ T        cells with an IC50 of 1 nM or less.    -   37. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein is a        reversible dimer.    -   38. The IL-7 binding protein according to any one of the        preceding embodiments, wherein the IL-7 binding protein inhibits        signaling, activation, cytokine production and proliferation of        CD4+ T cells and/or CD8+ T cells.    -   39. A nucleic acid encoding the IL-7 binding protein according        to any one of the preceding embodiments.    -   40. The nucleic acid of embodiment 39, wherein the nucleic acid        comprises a sequence having at least 80% identity to the nucleic        acid sequence set out in SEQ ID NO:13 encoding the light chain.    -   41. The nucleic acid of embodiment 39 or 40, wherein the nucleic        acid comprises the nucleic acid sequence set out in SEQ ID NO:13        encoding the light chain.    -   42. The nucleic acid of embodiment any one of embodiments 39-41,        wherein the nucleic acid comprises a sequence having at least        80% identity to the nucleic acid sequence set out in SEQ ID        NO:14 encoding the heavy chain.    -   43. The nucleic acid of any one of embodiments 39-42, wherein        the nucleic acid comprises the nucleic acid sequence set out in        SEQ ID NO:14 encoding the heavy chain.    -   44. The nucleic acid of any one of embodiments 39-43, wherein        the nucleic acid further comprises a sequence having at least        80% identity to the nucleic acid sequence set out in SEQ ID        NO:15 encoding a signal peptide.    -   45. A vector comprising a nucleic acid according to any one of        embodiments 39-44.    -   46. The vector of embodiment 45, further comprising a promoter        functional in a mammalian cell.    -   47. A host cell comprising the nucleic acid according to any one        of embodiments 39-44, or the vector according to embodiment 45        or 46.    -   48. A host cell according to embodiment 47, wherein the host        cell is a CHO cell.    -   49. A method of making the IL-7 binding protein according to any        one of embodiments 1-38, the method comprising maintaining the        host cell according to embodiment 47 or 48 in a medium to        produce the IL-7 binding protein and isolating or purifying the        IL-7 binding protein produced by the host cell.    -   50. An IL-7 binding protein produced by the method of embodiment        49.    -   51. An IL-7 binding protein that competes for binding to IL-7        with the IL-7 binding protein of any one of embodiments 1-38.    -   52. A pharmaceutical composition, comprising a        pharmaceutically-acceptable carrier and an IL-7 binding protein        or an IL-7 binding fragment thereof that exhibits binding for        IL-7 at an epitope comprising at least 5 contiguous amino acids        of a sequence set out in SEQ ID NO:12.    -   53. The pharmaceutical composition of embodiment 52, wherein the        IL-7 binding protein is according to any one of embodiments        1-38.    -   54. A pharmaceutical composition comprising the IL-7 binding        protein according to any one of embodiments 1-38 and a        pharmaceutically acceptable carrier or excipient.    -   55. The pharmaceutical composition according to any one of        embodiments 52-54, wherein the pharmaceutical composition        further comprises at least one additional therapeutic agent.    -   56. The pharmaceutical composition according to any one of        embodiments 52-55, wherein the pharmaceutical composition has a        pH of 4.5-7.0.    -   57. The pharmaceutical composition according to any one of        embodiments 52-56, wherein the pharmaceutical composition has a        pH of 5.5, 6.0, 6.2 or 6.5.    -   58. The IL-7 binding protein according to any one of embodiments        1-38, for use in therapy.    -   59. A method for treatment of an autoimmune and/or inflammatory        condition in a subject in need thereof, comprising administering        to the subject a therapeutically effective amount of the IL-7        binding protein according to any one of embodiments 1-38 or the        pharmaceutical composition of embodiment 52-57.    -   60. The method according to embodiment 59, wherein the        administering is transarterially, subcutaneously, intradermally,        intratumorally, intranodally, intramedullary, intramuscularly,        intravenously or intraperitoneally.    -   61. The method according to embodiment 60, wherein the        administering is subcutaneously.    -   62. The method according to any one of embodiments 59-61,        wherein the therapeutically effective amount is at least about        0.1 mg/kg, 0.5 mg/kg, 1 mg/kg, 3 mg/kg, 5 mg/kg, 10 mg/kg, 20        mg/kg, 30 mg/kg, or 40 mg/kg.    -   63. The method according to any one of embodiments 59-62,        wherein the therapeutically effective amount is about 3 mg/kg.    -   64. The method according to any one of embodiments 59-63,        wherein the therapeutically effective amount is administered to        the subject at least about once every 1-60 days.    -   65. The method according to any one of embodiments 59-64,        wherein the therapeutically effective amount is administered to        the subject once every 4 weeks.    -   66. The method according to any one of embodiments 59-65,        wherein the autoimmune and/or inflammatory condition is        Sjögren's syndrome.    -   67. The method according to any one of embodiments 59-65,        wherein the autoimmune and/or inflammatory condition is        rheumatoid arthritis.    -   68. The method according to any one of embodiments 59-65,        wherein the autoimmune and/or inflammatory condition is multiple        sclerosis.    -   69. The method according to embodiment 68, wherein the multiple        sclerosis is clinically isolated syndrome, relapsing-remitting,        primary progressive or secondary progressive.    -   70. The method according to any one of embodiments 59-65,        wherein the autoimmune and/or inflammatory condition is Crohn's        disease.    -   71. The method according to any one of embodiments 59-65,        wherein the autoimmune and/or inflammatory condition is        ulcerative colitis.    -   72. The method according to any one of embodiments 59-65,        wherein the autoimmune and/or inflammatory condition is lupus        erythematosus.    -   73. The use of the IL-7 binding protein according to any one of        embodiments 1-38, in the manufacture of a medicament for        treatment of an autoimmune and/or inflammatory condition.    -   74. The use according to embodiment 73, wherein the autoimmune        and/or inflammatory condition is Sjögren's syndrome.    -   75. The use according to embodiment 73, wherein the autoimmune        and/or inflammatory condition is rheumatoid arthritis.    -   76. The use according to embodiment 73, wherein the autoimmune        and/or inflammatory condition is multiple sclerosis.    -   77. The use according to embodiment 76, wherein the multiple        sclerosis is clinically isolated syndrome, relapsing remitting,        primary progressive or secondary progressive.    -   78. The use according to embodiment 73, wherein the autoimmune        and/or inflammatory condition is Crohn's disease.    -   79. The use according to embodiment 73, wherein the autoimmune        and/or inflammatory condition is ulcerative colitis.    -   80. The use according to embodiment 73, wherein the autoimmune        and/or inflammatory condition is lupus erythematosus.    -   81. An IL-7 binding protein of any one of embodiments 1-38, for        use in treatment of an autoimmune and/or inflammatory condition.    -   82. The IL-7 binding protein for use according to embodiment 81,        wherein the autoimmune and/or inflammatory condition is        Sjögren's syndrome.    -   83. The IL-7 binding protein for use according to embodiment 81,        wherein the autoimmune and/or inflammatory condition is        rheumatoid arthritis.    -   84. The IL-7 binding protein for use according to embodiment 81,        wherein the autoimmune and/or inflammatory condition is multiple        sclerosis.    -   85. The IL-7 binding protein for use according to embodiment 84,        wherein the multiple sclerosis is clinically isolated syndrome,        relapsing remitting, primary progressive or secondary        progressive.    -   86. The IL-7 binding protein for use according to embodiment 81,        wherein the autoimmune and/or inflammatory condition is Crohn's        disease.    -   87. The IL-7 binding protein for use according to embodiment 81,        wherein the autoimmune and/or inflammatory condition is        ulcerative colitis.    -   88. The IL-7 binding protein for use according to embodiment 81,        wherein the autoimmune and/or inflammatory condition is lupus        erythematosus.    -   89. Use of the IL-7 binding protein according to any one of        embodiments 1-38 or 50-51, for diagnosis of a disease or        condition.    -   90. A composition comprising the IL-7 binding protein according        to any one of embodiments 1-38 or 50-51, bound to a moiety or an        antigenic fragment thereof.    -   91. The composition of embodiment 90, wherein the moiety is IL-7        or a fragment thereof.    -   92. A solid support comprising the IL-7 binding protein        according to any one of embodiments 1-38 or 50-51.    -   93. The solid support of embodiment 92, wherein the solid        support is an array. 94. A device that comprises:    -   a. the solid support according to embodiment 92 or 93, and    -   b. a processor for detecting a signal, wherein the signal is        indicative of a binding of a moiety to the IL-7 binding protein        according to any one of embodiments 1-38 or 50-51.    -   95. A pre-filled syringe or autoinjector device, comprising the        IL-7 binding protein according to any one of embodiments 1-38,        50-51 or the pharmaceutical composition according to any one of        embodiments 52-57.    -   96. A kit comprising the IL-7 binding protein according to any        one of embodiments 1-38 or 50-51 and instructions for use.    -   97. An IL-7 binding protein comprising a light chain having at        least 80% identity to the amino acid sequence set out in SEQ ID        NO:18, SEQ ID NO:20, or SEQ ID NO:22.    -   98. An IL-7 binding protein of embodiment 97, comprising at        least 80% identity to a CDR of SEQ ID NO:18, SEQ ID NO:20, or        SEQ ID NO:22.    -   99. An IL-7 binding protein comprising a heavy chain having at        least 80% identity to the amino acid sequence set out in SEQ ID        NO:19, SEQ ID NO:21, or SEQ ID NO:23.    -   100. An IL-7 binding protein of embodiment 99, comprising at        least 80% identity to a CDR of SEQ ID NO:19, SEQ ID NO:21, or        SEQ ID NO:23.    -   101. An IL-7 binding protein comprising a light chain having the        amino acid sequence set out in SEQ ID NO:18, SEQ ID NO:20, or        SEQ ID NO:22 and a heavy chain having the amino acid sequence        set out in SEQ ID NO:19, SEQ ID NO:21, or SEQ ID NO:23.    -   102. An IL-7 binding protein comprising a CDR of SEQ ID NO:18,        SEQ ID NO:20, or SEQ ID NO:22 and a CDR of SEQ ID NO:19, SEQ ID        NO:21, or SEQ ID NO:23.    -   103. An IL-7 binding protein comprising a light chain having the        amino acid sequence set out in SEQ ID NO:18 and a heavy chain        having the amino acid sequence set out in SEQ ID NO:19.    -   104. An IL-7 binding protein comprising a light chain having the        amino acid sequence set out in SEQ ID NO:20 and a heavy chain        having the amino acid sequence set out in SEQ ID NO:21.    -   105. An IL-7 binding protein comprising a light chain having the        amino acid sequence set out in SEQ ID NO:22 and a heavy chain        having the amino acid sequence set out in SEQ ID NO:23.    -   106. A composition comprising the IL-7 binding protein of any        one of embodiments 97-105.    -   107. An IL-7 binding protein of any one of embodiments 97-105,        for use in treatment of an autoimmune and/or inflammatory        condition, wherein the autoimmune and/or inflammatory condition        comprises Sjögren's syndrome, rheumatoid arthritis, multiple        sclerosis (clinically isolated syndrome, relapsing remitting,        primary progressive or secondary progressive), Crohn's disease,        ulcerative colitis, or lupus erythematosus.    -   108. A method for treatment of an autoimmune and/or inflammatory        condition comprising administering to a subject the composition        of embodiment 106.    -   109. The method of embodiment 108, wherein the autoimmune and/or        inflammatory condition comprises Sjögren's syndrome, rheumatoid        arthritis, multiple sclerosis (clinically isolated syndrome,        relapsing remitting, primary progressive or secondary        progressive), Crohn's disease, ulcerative colitis, or lupus        erythematosus.    -   110. The use of the IL-7 binding protein according to any one of        embodiments 97-105, in the manufacture of a medicament for        treatment of an autoimmune and/or inflammatory condition.    -   111. The use of embodiment 110, wherein the autoimmune and/or        inflammatory condition comprises Sjögren's syndrome, rheumatoid        arthritis, multiple sclerosis (clinically isolated syndrome,        relapsing remitting, primary progressive or secondary        progressive), Crohn's disease, ulcerative colitis, or lupus        erythematosus.    -   112. A kit comprising the IL-7 binding protein according to any        one of embodiments 97-105, and instructions for use.

Sequences SEQ ID NO 1: human IL-7 Sequence with leader sequenceMFHVSFRYIFGLPPLILVLLPVASSDCDIEGKDGKQYESVLMVSIDQLLDSMKEIGSNCLNNEFNFFKRHICDANKEGMFLFRAARKLRQFLKMNSTGDFDLHLLKVSEGTTILLNCTGQVKGRKPAALGEAQPTKSLEENKSLKEQKKLNDLCFLKRLLQEIKTCWNKILMGTKEH SEQ ID NO 2: DRSPAI-L7B heavy chainQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGVHWVRQAPGKGLEWLAAIWTGGSTDYNSAFSSRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNGYGESFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO 3: DRSPAI-L7B light chainDIQMTQSPSSLSASVGDRVTITCKASESLDHDGDSYINWYQQKPGKAPKLLIYMGSNVEFGVPARFSGSGSGTDFTLTISSLQPEDFATYYCQQSNVDPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO 4: DRSPAI-L7B V_(H)QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGVHWVRQAPGKGLEWLAAIWTGGSTDYNSAFSSRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNGYGESFAYWGQGTLVTVSS SEQ ID NO 5: DRSPAI-L7B V_(L)DIQMTQSPSSLSASVGDRVTITCKASESLDHDGDSYINWYQQKPGKAPKLLIYMGSNVEFGVPARFSGSGSGTDFTLTISSLQPEDFATYYCQQSNVDPLTFGGGTKVEIK SEQ ID NO 6: DRSPAI-L7B CDRH1 SYGVHSEQ ID NO 7: DRSPAI-L7B CDRH2 AIWTGGSTDYNSAFSSSEQ ID NO 8: DRSPAI-L7B CDRH3 NGYGESFAY SEQ ID NO 9: DRSPAI-L7B CDRL1KASESLDHDGDSYIN SEQ ID NO 10: DRSPAI-L7B CDRL2 MGSNVEFSEQ ID NO 11: DRSPAI-L7B CDRL3 QQSNVDPLTSEQ ID NO 12: DRSPAI-L7B 1^(st) protected site FKRHICDANKEGMFLSEQ ID NO 13: Nucleic acid sequence encoding DRSPAI-L7B light chainGACATCCAGATGACCCAGAGCCCTAGCAGCCTGAGCGCCAGCGTGGGAGACAGGGTGACCATCACCTGCAAGGCCAGCGAGTCCCTGGACCACGACGGCGACAGCTACATCAACTGGTACCAGCAGAAGCCCGGCAAGGCCCCCAAGCTGCTGATCTACATGGGCAGCAACGTGGAGTTCGGCGTGCCCGCCAGGTTTAGCGGCAGCGGCAGCGGCACCGACTTCACCCTGACCATCAGCAGCCTCCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGAGCAACGTGGACCCCCTGACTTTCGGCGGCGGCACCAAGGTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO 14: Nucleic acid sequence encoding DRSPAI-L7B heavy chainCAGGTGCAGCTGGTGGAGAGCGGCGGCGGCGTGGTCCAGCCCGGAAGGAGCCTGAGGCTGAGCTGCGCCGCCAGCGGCTTCACCTTCAGCAGCTACGGGGTCCACTGGGTGAGGCAGGCCCCCGGAAAGGGCCTGGAGTGGCTGGCCGCCATCTGGACCGGCGGCTCCACCGACTACAACAGCGCCTTCAGCAGCAGGTTCACCATCAGCAGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACAGCCTCAGGGCCGAGGACACCGCCGTGTACTATTGCGCAAGGAACGGCTACGGCGAGAGCTTCGCCTACTGGGGCCAGGGCACCCTGGTGACCGTGAGCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGGCCGGAGCCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO 15: Nucleic acid sequence encoding signaling peptideATGGGCTGGTCCTGCATCATCCTGTTTCTGGTGGCCACCGCCACCGGTGTGCACAGCSEQ ID NO: 16: DRSPAI-L7B 2^(nd) protected site FKRHICDANKEGMFSEQ ID NO 17: Human IL-7 Sequence (without secretory leader)DCDIEGKDGKQYESVLMVSIDQLLDSMKEIGSNCLNNEFNFFKRHICDANKEGMFLFRAARKLRQFLKMNSTGDFDLHLLKVSEGTTILLNCTGQVKGRKPAALGEAQPTKSLEENKSLKEQKKLNDLCFLKRLLQEIKTCWNKILMGTKEHSEQ ID NO 18: A1290 light chainDIQMTQSPSSLSASVGDRVTITCKASQSVDDDGDSFINWYQQKPGKAPKLLIYVASNLESGVPARFSGSGSGTDFTLTISSLQPEDFATYYCQQSNEDPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO 19: A1290 heavy chainQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGLHWVRQAPGKGLEWLAAIWTGGSTDYNAAFISRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNGYGESFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO 20: A1291 light chainDIQMTQSPSSLSASVGDRVTITCKASHSVDDDGDSYMNWYQQKPGKAPKLLIYMASNLESGVPARFSGSGSGTDFTLTISSLQPEDFATYYCQQSNEDPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO 21: A1291 heavy chainQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGLHWVRQAPGKGLEWLAAIWTGGSTDYNAEFSSRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNGYGESFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO 22: A1294 light chainDIQMTQSPSSLSASVGDRVTITCKASQSVDDDGDSYMNWYQQKPGKAPKLLIYMASNLESGVPARFSGSGSGTDFTLTISSLQPEDFATYYCQQSNEDPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO 23: A1294 heavy chainQVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGVHWVRQAPGKGLEWLAAIWSGGSTDYNVAFSSRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNGYGESFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO 24: A1290 V_(L)DIQMTQSPSSLSASVGDRVTITCKASQSVDDDGDSFINWYQQKPGKAPKLLIYVASNLESGVPARFSGSGSGTDFTLTISSLQPEDFATYYCQQSNEDPLTFGGGTKVEIK SEQ ID NO 25: A1290 V_(H)QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGLHWVRQAPGKGLEWLAAIWTGGSTDYNAAFISRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNGYGESFAYWGQGTLVTVSS SEQ ID NO 26: A1290 CDRH1 SYGLHSEQ ID NO 27: A1290 CDRH2 AIWTGGSTDYNAAFIS SEQ ID NO 28: A1290 CDRH3NGYGESFAY SEQ ID NO 29: A1290 CDRL1 KASQSVDDDGDSFINSEQ ID NO 30: A1290 CDRL2 VASNLES SEQ ID NO 31: A1290 CDRL3 QQSNEDPLTSEQ ID NO 32: A1291 V_(L)DIQMTQSPSSLSASVGDRVTITCKASHSVDDDGDSYMNWYQQKPGKAPKLLIYMASNLESGVPARFSGSGSGTDFTLTISSLQPEDFATYYCQQSNEDPLTFGGGTKVEIK SEQ ID NO 33: A1291 V_(H)QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGLHWVRQAPGKGLEWLAAIWTGGSTDYNAEFSSRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNGYGESFAYWGQGTLVTVSS SEQ ID NO 34: A1291 CDRH1 SYGLHSEQ ID NO 35: A1291 CDRH2 AIWTGGSTDYNAEFSS SEQ ID NO 36: A1291 CDRH3NGYGESFAY SEQ ID NO 37: A1291 CDRL1 KASHSVDDDGDSYMNSEQ ID NO 38: A1291 CDRL2 MASNLES SEQ ID NO 39: A1291 CDRL3 QQSNEDPLTSEQ ID NO 40: A1294 V_(L)DIQMTQSPSSLSASVGDRVTITCKASQSVDDDGDSYMNWYQQKPGKAPKLLIYMASNLESGVPARFSGSGSGTDFTLTISSLQPEDFATYYCQQSNEDPLTFGGGTKVEIK SEQ ID NO 41: A1294 V_(H)QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGVHWVRQAPGKGLEWLAAIWSGGSTDYNVAFSSRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNGYGESFAYWGQGTLVTVSS SEQ ID NO 42: A1294 CDRH1 TYGVHSEQ ID NO 43: A1294 CDRH2 AIWSGGSTDYNVAFSS SEQ ID NO 44: A1294 CDRH3NGYGESFAY SEQ ID NO 45: A1294 CDRL1 KASQSVDDDGDSYMNSEQ ID NO 46: A1294 CDRL2 MASNLES SEQ ID NO 47: A1294 CDRL3 QQSNEDPLT

1.-35. (canceled)
 36. A method for the treatment of an autoimmune and/orinflammatory condition in a subject in need thereof, the methodcomprising administering to the subject a therapeutically effectiveamount of an interleukin 7 (IL-7) binding protein or IL-7 bindingfragment thereof comprising: (i) a heavy chain variable region CDR1comprising the amino acid sequence of SEQ ID NO: 6; (ii) a heavy chainvariable region CDR2 comprising the amino acid sequence of SEQ ID NO: 7;(iii) a heavy chain variable region CDR3 comprising the amino acidsequence of SEQ ID NO: 8; (iv) a light chain variable region CDR1comprising the amino acid sequence of SEQ ID NO: 9; (v) a light chainvariable region CDR2 comprising the amino acid sequence of SEQ ID NO:10; and (vi) a light chain variable region CDR3 comprising the aminoacid sequence of SEQ ID NO:
 11. 37. The method of claim 36, wherein theIL-7 binding protein or IL-7 binding fragment thereof comprises a heavychain variable region having a sequence at least 90% identical to theamino acid sequence of SEQ ID NO: 4 and a light chain variable regionhaving a sequence at least 90% identical to the amino acid sequence ofSEQ ID NO:
 5. 38. The method of claim 37, wherein the IL-7 bindingprotein or IL-7 binding fragment thereof comprises a heavy chainvariable region comprising the amino acid sequence of SEQ ID NO: 4 and alight chain variable region comprising the amino acid sequence of SEQ IDNO:
 5. 39. The method of claim 36, wherein the IL-7 binding protein orIL-7 binding fragment thereof comprises a heavy chain having a sequenceat least 80% identical to the amino acid sequence of SEQ ID NO: 2 and alight chain having a sequence at least 80% identical to the amino acidsequence of SEQ ID NO:
 3. 40. The method of claim 39, wherein the IL-7binding protein or IL-7 binding fragment thereof comprises a heavy chaincomprising the amino acid sequence of SEQ ID NO: 2 and a light chaincomprising the amino acid sequence of SEQ ID NO:
 3. 41. The method ofclaim 36, wherein the IL-7 binding protein or IL-7 binding fragmentthereof is an antibody.
 42. The method of claim 41, wherein the antibodycomprises an IgG1 or IgG4 Fc region.
 43. The method of claim 42, whereinthe antibody comprises a human IgG1 heavy chain constant region.
 44. Themethod of claim 43, wherein the antibody comprises a human IgG1 heavychain constant region having an alanine residue at position 235 andposition 237 according to EU numbering.
 45. The method of claim 36,wherein the autoimmune and/or inflammatory condition is multiplesclerosis, Sjögren's syndrome, rheumatoid arthritis, inflammatory boweldisease, Crohn's disease, ulcerative colitis, systemic lupuserythematosus, or type I diabetes.
 46. The method of claim 45, whereinthe multiple sclerosis is clinically isolated syndrome, relapsedremitting, primary progressive or secondary progressive.
 47. The methodof claim 45, wherein the autoimmune and/or inflammatory condition issystemic lupus erythematosus.
 48. The method of claim 45, wherein theautoimmune and/or inflammatory condition is inflammatory bowel disease.49. The method of claim 45, wherein the autoimmune and/or inflammatorycondition is type 1 diabetes.
 50. The method of claim 38, wherein theautoimmune and/or inflammatory condition is multiple sclerosis,Sjögren's syndrome, rheumatoid arthritis, inflammatory bowel disease,Crohn's disease, ulcerative colitis, systemic lupus erythematosus, ortype I diabetes.
 51. The method of claim 50, wherein the multiplesclerosis is clinically isolated syndrome, relapsed remitting, primaryprogressive or secondary progressive.
 52. The method of claim 50,wherein the autoimmune and/or inflammatory condition is systemic lupuserythematosus, inflammatory bowel disease, or type I diabetes.
 53. Themethod of claim 40, wherein the autoimmune and/or inflammatory conditionis multiple sclerosis, Sjögren's syndrome, rheumatoid arthritis,inflammatory bowel disease, Crohn's disease, ulcerative colitis,systemic lupus erythematosus, or type I diabetes.
 54. The method ofclaim 53, wherein the multiple sclerosis is clinically isolatedsyndrome, relapsed remitting, primary progressive or secondaryprogressive.
 55. The method of claim 53, wherein the autoimmune and/orinflammatory condition is systemic lupus erythematosus, inflammatorybowel disease, or type I diabetes.